COULTER LH 500 HEMATOLOGY ANALYZER

Transcription

1 COULTER LH 500 HEMATOLOGY ANALYZER TRAINING MODULES

2 WARNINGS AND PRECAUTIONS READ ALL PRODUCT MANUALS AND CONSULT WITH BECKMAN COULTER-TRAINED PERSONNEL BEFORE ATTEMPTING TO OPERATE INSTRUMENT. DO NOT ATTEMPT TO PERFORM ANY PROCEDURE BEFORE CAREFULLY READING ALL INSTRUCTIONS. ALWAYS FOLLOW PRODUCT LABELING AND MANUFACTURER S RECOMMENDATIONS. IF IN DOUBT AS TO HOW TO PROCEED IN ANY SITUATION, CONTACT YOUR BECKMAN COULTER REPRESENTATIVE. HAZARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS WARNINGS, CAUTIONS and IMPORTANTS alert you as follows: WARNING Can cause injury. CAUTION Can cause damage to the instrument. IMPORTANT Can cause misleading results. BECKMAN COULTER, INC. URGES ITS CUSTOMERS TO COMPLY WITH ALL NATIONAL HEALTH AND SAFETY STANDARDS SUCH AS THE USE OF BARRIER PROTECTION. THIS MAY INCLUDE, BUT IT IS NOT LIMITED TO, PROTECTIVE EYEWEAR, GLOVES AND SUITABLE LABORATORY ATTIRE WHEN OPERATING OR MAINTAINING THIS OR ANY OTHER AUTOMATED LABORATORY ANALYZER. WARNING Risk of operator injury if: All doors, covers and panels are not closed and secured in place prior to and during instrument operation. The integrity of safety interlocks and sensors is compromised. Instrument alarms and error messages are not acknowledged and acted upon. You contact moving parts. You mishandle broken parts. Doors, covers and panels are not opened, closed, removed and/or replaced with care. Improper tools are used for troubleshooting. To avoid injury: Keep doors, covers and panels closed and secured in place while the instrument is in use. Take full advantage of the safety features of the instrument. Do not defeat safety interlocks and sensors. Acknowledge and act upon instrument alarms and error messages. Keep away from moving parts. Report any broken parts to your Beckman Coulter Representative. Open/remove and close/replace doors, covers and panels with care. Use the proper tools for troubleshooting. CAUTION System integrity might be compromised and operational failures might occur if: This equipment is used in a manner other than specified. Operate the instrument as instructed in the Product Manuals and the on-line Help system. You introduce software that is not authorized by Beckman Coulter into your computer. Only operate your system s computer with software authorized by Beckman Coulter. You install software that is not an original copyrighted version. Only use software that is an original copyrighted version to prevent virus contamination. IMPORTANT If you purchased this product from anyone other than Beckman Coulter or an authorized Beckman Coulter distributor, and, if it is not presently under a Beckman Coulter service maintenance agreement, Beckman Coulter cannot guarantee that the product is fitted with the most current mandatory engineering revisions or that you will receive the most current information bulletins concerning the product. If you purchased this product from a third party and would like further information concerning this topic, call your Beckman Coulter Representative. ii of viii

3 TRADEMARKS 5C, COULTER, COULTER CLENZ, COULTER COUNTER, ISOTON, LIN-C, LYSE S, S- CAL AND ZAPOGLOBIN are trademarks of Beckman Coulter, Inc. All other trademarks, service marks, products or services are trademarks or registered trademarks of their respective holders. Copyright Beckman Coulter, Inc All Rights Reserved iii of viii

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5 REVISION STATUS Revision A Initial Issue (October 2003) Software Release 1A Revision B (April 2005) Software Release 2A Revision C (November 2007) Software Release 2A2 Revision CA (September 2009) Software Release 2A4 v of viii

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7 TABLE OF CONTENTS COURSE GUIDE PREREQUISITE TOPICS TH Theory REQUIRED TOPICS CAL Calibration HL History Logs QC Running Controls QC2 Additional QC RFC Replacing Components RPS Run Samples RSR Reagent Setup SC1 Setup Controls 1 SD Shutdown SF Sample Flow SU Startup TB Troubleshooting Basics WB Workstation Basics ADVANCED TOPICS ITT Introduction to Truth Table MT Mentor/Trainer LAMINATED SUMMARY SHEETS (back pocket) COULTER LH 500 TRAINING MODULES vii of viii

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9 COURSE GUIDE TABLE OF CONTENTS Course Map and Personal Progress Summary for COULTER LH Group Discussions Reminders How This Course Works Summary of Module Objectives of Coulter

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11 COULTER LH 500 HEMATOLOGY ANALYZER COULTER LH 500 TRAINING MODULES COURSE MAP Advanced Topics ITT Introduction to Truth Tables MT Mentor/Trainer Required Topics SD* Shutdown RPS* Run Samples CAL Calibration HL History Logs QC2 Additional QC SC1 Setup Controls 1 TB Troubleshooting Basics RC Replace Components QC* Running Controls RSR Reagent Setup SF Sample Flow SU* Startup WB Workstation Basics Prerequisite Topics * Daily Procedure TH Theory 3of 14

12 PERSONAL PROGRESS SUMMARY Name Prerequisite Topics Date Achieved Sign-Off Apps TH Theory Required Topics CAL Calibration HL History Logs QC* Running Controls QC2 Additional QC RFC Replace Components RPS* Run Samples RSR Reagent Setup SC1 Setup Controls 1 SD* Shutdown SF Sample Flow SU* Startup TB Troubleshooting Basics WB Workstation Basics Advanced Topics ITT Introduction to Truth Tables MT Mentor Trainer * Daily Procedures 4 of 14

13 GROUP DISCUSSIONS The following discussions take place in the classroom/laboratory. Ending times noted are approximate. Monday 8:15-11:00 General Information Learn about each other First day self-evaluation Safety: Personal Protective Equipment and Fire Evacuation Introduction to the course and materials Location of additional reference materials Instrument Demo Tuesday 8:15-9:30 Review of Patient Result Screens area Review of QA screens area Wednesday 8:15-10:00 Review PROService TM /RCS Info Flags, Codes and Messages Thursday 1:45-2:15 REMINDERS Last day self-evaluation, course evaluation and instructor evaluation Graduation You may take two fifteen minute breaks daily, one in the morning and one in the afternoon. There will be 45 minutes for lunch. Tuesday through Thursday, remember to complete the four daily procedures: Startup, Running and Reviewing Controls, Run Samples in the AM and Clean at the end of the day (about 15 minutes before leaving for the day). Each day when you arrive will begin with time to answer any questions you may have and to make any neccesary announcements for the day. 5of 14

14 How This Course Works This course is probably unlike any other training you may have experienced, unless you have taken a criterion-referenced course before. What is criterionreferenced instruction? Simply put, it is the learning of pre-specified performance criteria, using Skill Checks to verify your learning. The module provides information and guidance so you may practice the skills required to operate the LH 500 until you feel confident with them. A Skill Check determines whether you have achieved the stated objectives of each individual module. Let s look at the makeup of this course. The main features of this training are: Reviewing information and skills you already know so that you can do Skill Checks right away. Learning only skills that are necessary for your job (however, make the best of your time while you are here so that it is most beneficial for you and your lab). Working at your own pace. Practice a new skill until you achieve competency. Course Map The course map shows how each module of the course relates to other modules and to the course as a whole. It is a blueprint of the course. This course has three course maps to choose from depending on your particular instrument configuration. To use a course map, start at the bottom and work your way up to the top. Modules Read the Objectives and What to Learn, at the beginning of a new module. Complete the Skill Check for the module if you already know the task. It is a good idea to read (overview) the module anyway to make sure there are no terminology or procedural surprises. Read the module and complete the practice exercises for new material presented in the module. Refer to additional resources provided for the module. Work through the module at your own pace. This is not a race. If you are working with a partner, be sure both have a chance to practice and understand the information and exercises. 6 of 14

15 Resources Each module lists the resources available to complete the topic. Consult any resource that is appropriate to your needs. You may use some or all of the resources. A Facilitator is always available to answer any questions you may have. Please use the signup sheet provided. Additional resources are available for your reference use. Your facilitator will show you where they are located. Feel free to refer to them at any time during the week. Some modules require you to work with a colleague or to have a colleague review your work against a checklist. You may work with colleagues at other times too, if you wish. (Note: Skill Checks must be your own work unless otherwise noted.) Skill Checks Each module has a criterion check called a Skill Check. You may complete a Skill Check when you feel ready. Before doing so, you will save yourself time if you first make sure you can answer yes to these questions: Did I practice the skill(s) called for in the objective? Did I get a colleague sign-off, if it is called for by the module? If, after reading the module, you feel ready to complete the Skill Check without further study, do so. If your performance on a Skill Check is inadequate, you may, after further study and at the facilitator s discretion, be asked to complete the same or a similar Skill Check. When you complete a Skill Check, a facilitator or colleague (as indicated in the sign-off box) will check your work. Personal Progress Summary Ask whomever is certifying your work (course facilitator or colleague) to date and initial your Personal Progress Summary next to the appropriate module. Master Progress Chart Facilitators use a Master Progress Chart to keep track of the progress of the entire class. Once certified on a module, be sure a facilitator makes the proper entry on the master progress chart. Until a facilitator makes this entry, they assume that you have not yet mastered the module. 7of 14

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17 SUMMARY OF MODULE OBJECTIVES PREREQUISITE TOPICS TH THEORY Given an operating COULTER LH 500 Workstation or another PC satisfying minimum requirements Complete Coulter Theory Topics, Chapters One through Five on the CD- ROM in the pre-requisite package. Print test scores OR record scores on the attached sheet and bring with you to the first day of class. CAL CALIBRATION REQUIRED TOPICS Given an operating COULTER LH 500 and access to HELP, Perform calibration of an LH 500 Analyzer using the COULTER S-CAL Calibration kit. Adjust calibration factors as required by the calibration results. Tell when you should calibrate your instrument. HL HISTORY LOGS Given an operating COULTER LH 500 Workstation, and access to HELP, perform these procedures: Access the History Logs from the Command Center. Add a comment to a History Log entry. Access detailed descriptions and corrective actions for messages. Print a History Log. Archive a History Log (Elective). QC QUALITY CONTROL Given an operating COULTER LH 500, package inserts from control products, COULTER LATRON PRIMER and CONTROL, COULTER 5C Cell Control and COULTER Retic-C Cell Control and access to HELP Describe proper storage and handling requirements for LATRON PRIMER AND CONTROL, 5C Cell Control and Retic-C Cell Control and perform QC. Analysis according to the package inserts and HELP procedures 9 of 14

18 Access HELP to find and use procedures for running LATRON, 5C Cell Control and Retic-C Cell Controls. Access control folders using the Control Tree. Process LATRON Primer and Control, 5C Cell Control, Retic-C Cell Control according to the package inserts and HELP procedure. Access HELP information for action to take when a control is outside expected ranges. Recognize flagged control results for any type of control run. Remove and restore control results. Add a comment to a control result. Use Levey-Jennings graphs to recognize trends and shifts. Identify the steps required to print a control result. Set up shift designations. QC2 Additional QC Given an operating COULTER LH 750 Workstation, access to HELP, and removable media Download control data for IQAP. Demonstrate how to set up and use lab limits. RC REPLACE COMPONENTS Given a COULTER LH 500 or a practice station requiring a component replacement, locate the procedure using HELP or use the procedure provided and perform the procedure following all safety precautions. RPS RUNNING PATIENT SAMPLES Given an operating COULTER LH 500, bloods collected in EDTA, other supplies, and access to HELP State the maximum aspiration volume needed for automatic and manual modes of operation. State the maximum stability of CBC/Diff and Retic samples stored at room temperature and stored refrigerated. Identify the two types of cassettes that can be used on the LH 500 and the proper tubes to use in each. Process bar coded samples in the automatic mode of operation and print results automatically. Process bar coded and non-bar coded samples in the manual mode of operation and print results automatically. Process a STAT sample. 10 of 14

19 Process samples in the Predilute mode. Identify the populations on the two-dimensional Dataplot for Diff and Retic. Find samples in the Database using Navigation buttons (Previous). RSR REAGENT SETUP AND REPLACEMENT Given a COULTER LH Workstation, access to HELP, Update the Reagent Setup screen in the LH Workstation (if necessary). Recognize instrument messages used for reagents. Properly change a reagent container. Properly replace a waste container. SC1 SETTING UP CONTROLS Given a COULTER LH 500, access to HELP, control package download instructions, removable media and control package assay sheet or Control diskette. Diskette will be phased out. Access the screen for setting up new control folders on the LH Workstation. Set up a COULTER LATRON Control folder using data from the package insert. Set up COULTER 5C Cell Control and COULTER Retic-C Cell using either the removable media, 2D Barcode Scanner or the Control diskette (to be phased out). Explain how a lab might use Auto Transmit and Auto Stop features. SD SHUTDOWN Given an operating COULTER LH 500: Perform the Clean procedure using COULTER CLENZ cleaning agent. State what cycle is performed at the end of the programmed Clean time. State the difference between the Clean procedure and the Shutdown procedure. State the minimum time for the LH 500 to be in Shutdown. State the maximum time for the LH 500 to be in Shutdown. State when it is necessary to perform Extended and Prolonged Shutdown. 11 of 14

20 SF SAMPLE FLOW Given an operating COULTER 500, Summary Sheet diagrams, blood sample, and access to HELP : Locate and name major Diluter components, using no references. List the constituents of WBC and RBC/Plt, dilutions, using references. State the four constants and one variable used for the COULTER Principle, using references. State the importance of sweep flow and mixing bubbles, using references. State the purpose of cross-rinse at the end of an instrument cycle, using references. SU STARTUP Given an operating COULTER LH 500 and access to HELP : Locate the STARTUP on the Command Center. Initiate Startup cycles on the LH 500 Analyzer. Access the Quality Assurance, Daily Checks windows to check results. Access the Daily Check Details window. Perform the proper procedures in the event of a message on the Daily Checks window, following the HELP procedures. Repeat Background from the Command Center. Suppress automatic printing of Daily Checks. Use (View History) button to view stored Daily Checks results. Return to the Daily Checks window using the View Current button. TB Troubleshooting Basics Given an operating LH 500, access to HELP, and access to the SF Sample Flow training module: Identify an instrument problem based on: Error messages. Abnormal sample results. Abnormal Daily check or System test results. Abnormal control results. Isolate a problem based on observation of sample flow. Isolate the component(s) involved based on observed symptoms. Perform selected cleaning and adjustment procedures as necessary. So that an operator can return an instrument to operating condition. 12 of 14

21 WB WORKSTATION BASICS Given a COULTER LH 500 Workstation and access to HELP, Tell whether a display is from the Instrument Computer or the Workstation describing to a facilitator and identifying the Instrument Computer and the Workstation by pointing to each Switch between the two computer displays using the switch box Use the conventions of the Windows 2000 Professional Edition Operating System using the mouse and keyboard Log on and log off the Workstation Identify the icons of the Command Center by labeling a picture Navigate through all Workstation screens using the mouse and/or keyboard following written instructions Identify Common and Specific Toolbars on various Workstation screens by pointing to them Use the on-line Help to find and print topic information and create favorites Shutdown and restart the Workstation to address error situations Tell how and why one uses radio buttons, check boxes, and Tool Tips Resize and Move Help windows Navigate through the Instrument Computer screen using the keyboard following written instructions Reboot Instrument Computer using the Standby/Reset switch Advanced Topics ITT INTRODUCTION TO TRUTH TABLES Given an operating COULTER LH 500, access to HELP, and this module Determine whether a specimen is clinically normal or abnormal (in your laboratory) Define review limits and review protocols (in your laboratory) Know criteria to follow for data collection Know how to interpret the Rumke Binomial Distribution Table Calculate the statistics necessary to determine acceptable performance Perform Truth Table Analysis Define the four categories on a Truth Table 13 of 14

22 MT MENTOR/TRAINER Given an operating LH 500, training modules, and appropriate supplies for administering modules: Format a training plan for your laboratory Verify skill performance for each trainee Have a course Facilitator and/or Applications Specialist verify your performance as a mentor/trainer 14 of 14

23 CAL CALIBRATION OBJECTIVES Given an operating COULTER LH 500, and access to HELP Perform calibration of an LH 500 Analyzer using the COULTER S-CAL Calibration Kit Adjust calibration factors as required by the calibration results Tell when you should calibrate your instrument Tell when you should verify calibration of your instrument Why Is It Important? Calibration fine-tunes the LH 500 Analyzer to give you the most accurate results possible. Resources To complete this module you will need LH 500 LH 500 Workstation with HELP One vial of EDTA blood One vial of COULTER S-CAL calibrator from the Calibration Kit (PN ) NOTE: The procedure in this module is adapted from HELP. Some steps may apply particularly to the classroom situation and not necessarily to your lab. CAL-1 of 10

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25 INFORMATION / PRACTICE SECTION COULTER LH 500 TRAINING MODULES The S-CAL calibration for CBC parameters must be performed by your laboratory after installation. Thereafter, you must follow your own laboratory, local or national regulations as to frequency of calibration. Sometimes a component involving dilution (such as a diluent dispenser) may need to be replaced. Your service engineer will perform a rough calibration afterward using your current controls to allow instrument operation to continue. However, an S-CAL verification should be performed within several days to a week to verify cal factors. Keep in mind that a new component should be given several days to break-in or settle before performing the verification. Calibrating CBC Parameters with S-CAL Calibrator Print the HELP topic Calibrating CBC Parameters with S-CAL Calibrator. You may wish to print the individual links for each step. NOTE: Room temperature should be stable and within the normal ambient temperature range. If the average ambient room temperature changes more than +/- 10 o F from the calibrating temperature, verify calibration and recalibrate (if necessary). Instrument Preparation Ensure Apertures Are Clean 1. a) If you routinely shutdown for at least 30 minutes with COULTER Clenz on a daily basis, and you are shutdown right now, other cleaning is unnecessary. b) If you routinely shutdown for at least 30 minutes with COULTER Clenz on a daily basis, but you are beginning calibration after processing patient samples, shutdown the instrument in the cleaning agent for 30 minutes before proceeding. (You could use the Clean Cycle if you wish). c) If you do not routinely shutdown for 30 minutes with COULTER Clenz on a daily basis, bleach the apertures before proceeding. Note: You will not need to do this in the classsroom. Verify Instrument Functions Properly 2. Check reagent containers for: Sufficient quantity: Diluent at least ½ full; LyseSIII - diff at least ¼ full Not beyond expiration date No precipitates, turbidity, particulate matter or unusual color Proper connections between the diluter and the reagent containers 3. Check the waste container (if you use one) for: Sufficient capacity Proper connections 4. Perform a Startup. (If you performed a Clean Cycle, startup occurs automatically.) CAL-3 of 10

26 Prepare S-CAL Calibrator 5. Take one of the prefilled S-CAL vials in the kit out of the refrigerator to allow it to equilibrate at room temperature for 15 minutes before using. (See package insert for details.) Prepare Instrument For Calibration 6. On the Workstation Command Center, ensure the blood detector is enabled (checked), the Process Type is set to Auto Analysis and the Run Type is set to C. Select Start. 7. Cycle a sample of normal whole blood as a prime. Select Stop. Obtaining S-Cal Reference Values o Get Removable Media for downloading S-CAL reference values from the Internet. o At your Individual Workstation, open the internet and access the following pathway to download calibrator reference values. Support>QA & Service Programs>Go to eiqap>hematology Assays o Follow the on-line instructions for downloading. o Once the S-CAL reference values are downloaded to the removable media, continue with the next steps. Calibration Set Up at the Workstation 8. On the Workstation Command Center, select then select (CBC Calibration). 9. Select CALIBRATION in the Process Type drop-down box. The background color of the Calibration window changes to the same background color as the Command Center. 10. If necessary, select (Clear table) to clear old calibration information from the database. The values on the window change to If you have already set up calibration information, check that the appropriate lot number for calibration is selected; otherwise, set up new calibration information. 12. Select (Calibration Setup) from the specific toolbar to display the Instrument Calibration Setup window. CAL-4 of 10

27 13. Select (Load S-CAL values). Insert your removable media into the LH 500 Workstation and follow the instructions on the pop-up window to load the reference values, lot number and expiration date. 14. Print the Instrument Calibration Setup window. Label it Pre-Cal. Save it for the Skill Check. 15. Select on the Instrument Calibration Setup window to save and close. 16. Take out the removable media. 17. Watch out for this relatively common error! On the CALIBRATION screen, check the Lot # box (on the left side of the screen) for the Lot # you just set up. If the Lot # is incorrect, use the drop-down box to select the correct Lot #. The Lot# is the 4 digit number on the S-CAL vial. NOTE: Calibration runs will be rejected if the calibrator information has not been set up, or if the calibrator is expired. If expired, the Expiration field will be backlighted RED. Run S-CAL Calibrator IMPORTANT Misleading results could occur if you fail to perform the calibration procedure within 1 hour of opening the S-CAL calibrator vials. Follow the instructions in the S-CAL calibrator package insert. 18. Prepare the prefilled S-CAL calibrator vial according to the instructions in the package insert. It is very important to follow the mixing directions carefully. 19. Set the number of aspirations to 11. (One vial of S-CAL is sufficient for 11 aspirations.) 20. Place the S-CAL vial into a cassette, and place the cassette in the right loading bay of the instrument. Select Start. 21. Automatic processing of the cassette begins. The Workstation automatically deletes the results from the first sample because the results are used as a prime. NOTE: The extra vial of calibrator in the kit may be used to calibrate another cap-pierce instrument or to repeat calibrator runs, if necessary. Review Results On The Calibration Screen On the CALIBRATION screen, inspect the calibration results table for trending. The parameter results must not show a trend. If the results show trending, there could be an instrument problem; call your Beckman Coulter representative. Do not continue. With the following in mind, use the calibration statistics to determine if you should transmit new calibration factors to the Analyzer. CAL-5 of 10

28 IMPORTANT Misleading results could occur if you transmit calibration factors that are outside the established limits. If results are outside the limits, call your Beckman Coulter representative. The Workstation checks the results for precision (%CV within the established limits; the same limits as used for Reproducibility.) The Workstation flags results outside the limits with a RED background (%CV, FAC % Diff and Delta Diff). They appear underlined on the printout. If you choose to transmit calibration factors other than those selected by the Workstation outside either the limits or factors that have verified a message appears indicating a Calibration Criteria Violation. You must confirm that you want to transmit the calibration factors. Once you confirm the transmission, the Workstation transmits the calibration factors and posts a message to the Calibration history log. IMPORTANT Misleading results could occur if you calibrate MCV when the RBC FAC% Diff is out of range because MCV depends on RBC. Do not calibrate MCV if the RBC FAC% Diff is out of range. The Workstation checks the FAC% Diff and DELTA Diff. The Workstation automatically selects the parameters that need adjustment and indicates results that meet the calibration criteria with a yellow background. If you do not want to adjust a marked parameter, unmark the check box. CALIBRATION CRITERIA TABLE Parameter Precision (CV%) Acceptable Fac%Diff Cal if Fac%Diff is Cal if Delta Diff is WBC CV<=2.5% <=5.0% >1.25% BUT<5.0% >0.1 BUT<=0.4 RBC CV<=2.0% <=2.0% >0.7% BUT<=2.0% >0.03 BUT<=0.09 Hgb CV<=1.5% <=3.0% >0.78% BUT<=3.0% >0.1 BUT<=0.4 MCV CV<=2.0% <=2.5% >1.18% BUT<=2.5% >1.0 BUT<=2.0% Plt CV<= 5.0% <=9.0% >2.70% BUT<=9.0% >6.0 BUT<=20.0 MPV CV<= 3.0% <=20.0% >5.0% BUT<=20.0% >0.5 BUT<=2.0 Are all parameters within limits? Are no boxes checked? If No, then no further action is necessary. Calibration is verified. If this is the case in class, be sure to contact a facilitator before proceeding. Are any boxes checked? If Yes, then parameter(s) need to be calibrated. CAL-6 of On the Workstation CALIBRATION screen, select (Adjust Calibration) on the Specific Toolbar. 23. The Adjust Calibration button is active only after the Workstation receives ten (10) valid calibration results. After calibration factors have been adjusted, the Adjust Calibration button is grayed out.

29 24. Select to print new calibration factors for your logbook. Print a copy of the Calibration Run screen and the now updated Instrument Calibration Setup screen (label it Post-Cal ) and save them for your Skill Check. After printing the Calibration Run Screen, make sure that you clear the screen to prevent recalculation of calibration factors that were changed. This happens when you exit this screen and come back to it at a later time. Don t do this until the skill check when the facilitator is present. 25. On the Command Center, set Process Type to Auto Analysis and set #aspirations back to Verify calibration factor changes by cycling each level of COULTER 5C Cell Control in the Automatic aspiration mode. Note: For this classroom exercise, it is not necessary to perform this verification. Be sure to do this in your own laboratory. Calibrate or Verify Calibration? 1. From the CALIBRATION screen, select on the Common Toolbar. 2. From the list on the left, select the topic: Calibration Overview. 3. Review the information about When to Calibrate to discuss as part of the Skill Check for this module. If you run S-CAL calibrator and change at least one parameter to a new cal factor, you have calibrated that parameter. Verify by running 5C Cell Controls. If you run S-CAL calibrator and make no changes to any cal factors, you have verified the current cal factors. No other verification is necessary. Proceed to the Skill Check for this module and bring the following with you: Calibration Set Up and Calibration Results screens printouts (three total) Calibration overview printout CAL-7 of 10

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31 CAL COULTER LH 500 TRAINING MODULES CALIBRATION SKILL CHECK 1. At your instrument, demonstrate to your facilitator how to automatically transmit calibration factors from the Workstation to the Analyzer. 2. Show the three calibration printouts (Calibration Run Table and two Calibration Set Up screens) to your facilitator. Answer the following questions. Circle the letter of the correct answer(s). 1. You should calibrate your COULTER LH 500: a. Monthly b. At installation c. After replacing the Blood Sampling Valve d. a and c are correct e. b and c are correct f. a and b are correct 2. Which of the following are valid reasons to run S-CAL calibrator to verify calibration settings? a. Controls show unusual trending b. On the Calibration Table a parameter checkbox is selected. c. Ambient room temperature changes by more than +/- 10 o F from calibrating temperature d. According to your laboratory or regulatory agency specifications 3. What parameters are calibrated on the LH 500 Analyzer? a. CBC and Diff b. CBC c. CBC, Diff and Retic d. CBC and Retic Facilitator sign off CAL-9 of 10

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33 HL HISTORY LOGS OBJECTIVES Given an operating COULTER LH 500 Workstation, and access to HELP, perform these procedures: Access the History Logs from the Command Center Add a comment to a History Log entry Edit a comment already in the History Log Print a History Log Archive a History Log (Elective) Why Is It Important? History Logs provide ongoing on-line documentation of instrument messages, reagent and control updating, as well as other information about your LH 500. History Logs eliminate the need for a handwritten Action Log, since all data can be printed and/or archived for permanent storage. Resources To complete this module you will need LH 500 Workstation with access to HELP Printer 3.5-inch diskette (if doing the archive elective) HL-1 of Coulter

34 OULTER LH 500 TRAINING MODULES INFORMATION / PRACTICE SECTION 1. At the Workstation from the Command Center, go to (History Logs) Each History Log has its own tab. The Event Log has general instrument messages. Other Logs such as Instrument, Daily Checks, Control Data, etc. contain more specific information. Messages are listed with the most recent first on the list. Practice Each log may have 2500 entries, then it rolls over (First in, First out). 2. From the History Logs window, select HELP. 3. Find and Print the topic, Working with Electronic History Logbooks, as well as the data from the hypertext delete some or all of the messages and add comments to a logbook. 4. Close HELP. Use the LH 500 Workstation at your instrument for the Practice part of this module. Adding a Comment 1. Select any History Log, using the tab. 2. Click on the date of the entry for which you want to make a comment. 3. Click on the (Add/Edit Comment) button. 4. Type a comment in the field provided (up to 256 characters). 5. Select. The comment is stored and displayed with the selected message. For the Skill Check, remember the message to which you added a comment so that you can show it to a facilitator. 6. Select a different comment (someone else s). Make an addition to it. Select HL-2 of 6

35 Printing from a History Log 1. Select any History Log. 2. Select the Print button. 3. Choose Selected Category. 4. Choose By Date: and enter the dates of a week ago. 5. Select. 6. Save the printout for the Skill Check. HL-3 of 6

36 OULTER LH 500 TRAINING MODULES Archiving (Elective) 1. Select any History Log. 2. Select the [Archive Result(s)] button. 3. Put a formatted 3.5-inch diskette into the A:\ drive of your LH 500 Workstation. 4. Select the radio button Selected Category. 5. Select By Date: 6. From your selected Log, type in a range of dates seen in the Log, for archiving. 7. Select. 8. The default filename for your archived file is the current date. NOTE: The default filename is, for example Select <SAVE> button. 10. Go to the computer with Microsoft Office and put the diskette with the archived file into the A:\ drive. 11. On the computer Desktop, click on the Excel icon on the MS Office toolbar. 12. On the Menu Bar, go to File Open. Under Look In, select 3½ Floppy (A:). Under Files of Type, select All Files. 13. Open the archived file from your diskette. 14. If another small window opens, follow the on-screen instructions to make the following selections (otherwise continue at step 15): a) Select Delimited. Select <Next>. b) Select TAB and COMMA. Select <Next>. c) Change date format, if desired. Select <Finish>. 15. In Excel, expand the column widths to fit the data in each field. 16. Go to Menu Bar, File Save As. 17. Choose Microsoft Excel Workbook (file type). Format is.xls. 18. Select Save. 19. Save the diskette for the Skill Check. If you can perform all the operations from the Practice part of the module, collect your diskette and printouts and proceed to the Skill Check. HL-4 of 6

37 HL HISTORY LOGS SKILL CHECK 1. Show a facilitator: A printed History Log of your choice. 2. At your instrument Workstation: Show a facilitator the History Log message to which you added a comment. 3. If you did the elective archiving: Using the computer having Microsoft Office, put the diskette with the archived file into the A:\ drive. Go to Microsoft Excel and access the archived History Log on the A:\ drive. Facilitator sign off HL-5 of Coulter

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39 QC QUALITY CONTROL OBJECTIVES Given an operating COULTER LH 500, the package inserts from control products, COULTER LATRON PRIMER and CONTROL, COULTER 5C Cell Control and COULTER Retic-C Cell Control and access to HELP Describe proper storage and handling requirements for LATRON PRIMER AND CONTROL, 5C Cell Control and Retic-C Cell Control and perform QC Analysis according to the package inserts and HELP procedures Access HELP to find and use procedures for running LATRON, 5C Cell Control and Retic-C Cell Controls Access control folders using the Control Tree Process LATRON Primer and Control, 5C Cell Control, Retic-C Cell Control according to the package inserts and HELP procedure Access HELP information for action to take when a control is outside expected ranges Recognize flagged control results for any type of control run Remove and restore control results Add a comment to a control result Use Levey-Jennings graphs to recognize trends and shifts Identify the steps required to print a control result Set up shift designations Why Is It Important? Understanding how to store, handle and use the control products available ensures reporting of the most accurate results possible to the clinician. Also, knowing how to review and output control results is an important follow-up to control processing. Resources NOTE: If you are coming from a non-coulter Hematology Analyzer, see a facilitator for a control processing demonstration. To complete this module you will need LH 500 LH Workstation with HELP Control package inserts for LATRON PRIMER and CONTROL, 5C Cell Control and Retic-C Cell Control Control products for processing Retic Prep kit (Elective) QC-1 of Coulter

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41 INFORMATION / PRACTICE SESSION Control Storage and Handling COULTER LH 500 TRAINING MODULES Refer to the control package inserts available in the lab to answer the following questions before you run controls. 1. LATRON Primer and Control are run at room temperature. TRUE FALSE 2. State the proper procedure for mixing LATRON Control. 3. 5C and Retic-C Cell Controls should be warmed at ambient temperature for minutes. 4. It s OK to mix 5C and/or Retic-C Cell Controls using a mechanical mixer. TRUE FALSE 5. You should return 5C and Retic-C Cell Controls to the refrigerator within minutes. 6. When you hand mix 5C Cell Control, how many 8 x 8 x 8 sets do you perform? When you hand mix Retic-C Cell Control, how many 8 x 8 x 8 sets do you perform? Show these answers to a facilitator as part of the Skill Check for this module. QC-3 of 12

42 Running LATRON Primer and Control Before running any controls go to (Run Configuration) and deselect the checkbox for Autoprint of controls, if it has been selected. Click on (OK) 1. At the Workstation Command Center select (from the Common Toolbar) You will be in Reviewing Control Results HELP topic. 2. Select Search, type in Run Control, (ensure Match similar words check box at the bottom of the screen is selected), click List Topics button. 3. Select and Display Running Latex (LATRON): Diff and Retic. 4. Print or read the procedure. 5. Close the HELP topic. NOTE: If you have never used LATRON Primer and Control, see a facilitator for additional details about using these products. 6. Run LATRON Primer and Control following the printed procedure. Watch out for these relatively common errors! When you run LATRON Primer, be sure to press Stop then choose CONTROL as the Process Type and Ltx Primer as the Run Type. Press Start.The aspiration mode defaults to Manual. The LH 500 compares the Primer results to the maximum value of 500. When you run LATRON Control, be sure to press Stop then choose Latex as the Run Type. Press Start. The LH 500 compares the results to those entered into the Workstation at control set up. QC-4 of 12

43 Latex Control Is Outside Expected Ranges 1. At the Workstation go to COULTER LH 500 TRAINING MODULES (from the Common Toolbar) Topic on the screen is: Reviewing Control Results 2. In the HELP window at the left of the screen, select the Contents tab, then LH 500 Analyzer Controls Click on and Display: When a Latex Control is Outside its Expected Ranges 3. Read the topic. You now have some basic troubleshooting tools to use if LATRON Control is outside its expected ranges. Running 5C Cell Control 1. In the HELP window at the left of the screen, click on: Cycling CBC/Diff Controls in the Automatic Mode. 2. Read the procedure. 3. Close HELP. Note: You do not have to be on the QA/QC screen when you run controls. 4. Use the HELP procedure to make the proper setup selections on the Command Center. 5. Place the properly mixed controls into cassette(s) with bar code labels facing up. 6. Place cassette(s) into the right-hand loading bay. 7. Use the Control Tree to access control folders and ensure that control results are in their correct folders. 8. Return the 5C Cell Control vials to the refrigerator. ABN II ABN I Norm Level I Level II Level III QC-5 of 12

44 Running Retic-C Cell Control (Elective) The Reticulocyte methodology on the LH 500 is semi-automated. It requires the use of the Retic Prep Kit (PN ) which contains two reagents, A and B. Reagent A modified New Methylene Blue stain provided in a dropper bottle Reagent B a clearing solution of very dilute sulfuric acid Look up and print the Help topic for running Retic-C Cell Control. The preparation steps must be followed exactly for accurate results: 1. Take control vials from refrigerator and prepare according to insert directions. 2. Label two 12 x 75 ml glass vials for each level of control 3. Add four drops of Reagent A into a labeled vial. 4. Pipette 50 µl of well-mixed control material and shake vial to mix control and stain. Allow to incubate at room temperature for a minimum of 5 minutes and a maximum of 60 minutes. 5. Prepare instrument to run the Retic-C control: Select Stop Choose Process Type CONTROL or AUTOANALYSIS, Run Type R. Select Start Be prepared to type the 6-digit control lot number 6. Gently remix the control/stain vial. Pipette 2 µl of the mixture into the bottom of the clean, labeled 12 x 75 ml glass vial for that control. 7. Immediately dispense 2 ml of Reagent B into the vial, holding the vial at an angle while doing so. This ensures complete mixing of the sample without forming bubbles. Do not shake. 8. Wait 30 seconds and then analyze the retic preparation you just made. Type in the 6-digit lot number in the Barcode field and press Enter or Tab Immerse aspirator tip in the prepared sample Press and release the sample bar (it will aspirate the entire contents) Remove the tube when you hear the beep 9. Review results. QC-6 of 12

45 5C and/or Retic-C Cell Controls are Outside Expected Ranges 1. At the Workstation QA/QC screen select (from the Common Toolbar). 2. In the HELP window at the left of the screen, select the Contents tab, then click on and Display the topic: When a Control is Outside its Expected Ranges. 3. Read the topic. You now have some basic troubleshooting tools to use if 5C Cell Control or Retic-C Cell Control is outside expected ranges. 4. Close HELP Finding Flagged Control Results 1. At the Workstation, from the Command Center, go to, Open the Control Tree by clicking on the in front of BCI. 2. Click on the the in front of 5C. All 5C folders open in numerical order. 3. Click directly on the 5C Cell Control folder you wish to open. The most recent result in the folder is at the top of the list. 4. Search through the 5C folders or Latron folder until you find a flagged result. Look for the Date and Time fields to have a red background. Use the scroll bar at the bottom of the results window to find the specific flagged results. Removing and Restoring a Control Result 1. Now that you ve found a flagged result, click on the deselect [Des] button at the left of the row. (If there are no flagged results, just use any run to practice.) The line is now grayed out or removed from the statistical results below. Note the total number of runs in the control folder after removing one run. 2. Practice removing and restoring control results, but leave at least one removed result in the folder for your final printout. QC-7 of 12

46 Add a Comment in a Control Folder 1. Choose a control run that you ran today. Click on the button. 2. Type a comment in the comment box and select [OK] to save the comment. 3. The Workstation adds the comment to the appropriate control sample and places an X in the CMNT column for the specific control. The comment is also added to the History Logs, Control Data tab. 4. Click on any box with an X in the CMNT column to view a comment. Note: When you print the file, the comment appears under the run it is associated with. 5. Select (History Logs) from the Command Center. Select the Control Data Tab and view the comment added to the control file. 6. Close the History Logs window. Levey-Jennings Graphs 1. From the Command Center select QA then QC and open a control folder. 2. Below the statistical information for a control folder are the thumbnail Levey- Jennings graphs showing the most recent 10 runs in the control folder. 3. The parameters are grouped in sets of three, starting with WBC, RBC, Hgb. The scroll bar at the top of the graphs lets you access graphs for other parameters. Scroll to the PLT tab and click on it now. Use the graphs to observe trends and shifts. 4. Double-click on any thumbnail graph to see the larger version Full Page graph. This graph shows up to 100 data points for the selected control folder. Do this now. Select [Close]. 5. You may print either version of the graphs. QC-8 of 12

47 Printing Control Results 1. With the control folder still open, on the Common Tool Bar select. 2. By selecting the desired radio buttons on the screen you can print various combinations of control folder data. 3. For this exercise, select to print Selected Lot and Thumbnail graphs. 4. After making your selection, click on. Setting Up and Using the Shift Clocks On the specific toolbar in the QA, QC screen area are a set of clock buttons. These allow you to review all the runs in a control file or to review by shift. Shift 0 is all runs from all shifts. Shifts 1, 2 or 3 are the runs by individual shift. Selecting any of these buttons automatically extracts the correct runs based on the shift time settings. The buttons are active only if shift times are entered.the shift time settings are entered in: System Setup Quality Assurance. QC-9 of 12

48 Practice 1. Select System Setup and 2. Select the tab. 3. Select Use Multiple Shifts. 4. View the area where shift times are entered. Please do not change what is setup on the your unit. Just cancel to back out. To practice, use an individual workstation (after this exercise) to enter the starting times of your shifts. Refer to the screens below. 5. Select (OK). 6. Go back to a control folder and print a set of data from just one shift. (Do Not print any Levey-Jennings graphs this time) 7. Save the printout for the Skill Check If you can discuss storage and handling of controls, process controls using HELP procedures, and discuss basic troubleshooting of controls that are outside expected ranges. Also, collect your printouts and perform the Skill Check for this module QC-10 of 12

49 QC QUALITY CONTROL SKILL CHECK Storage and Handling Answer the following questions: 1. LATRON Primer and Control may be stored at C or at room temperature. TRUE FALSE 2. COULTER 5C Cell Control should be mixed using which of the following techniques? a) using the LH Analyzer rocker bed b) by hand 8 x 8 x 8 two times c) using a rocker or rotating mixer d) any of the above 3. Storage of 5C Cell Control and Retic-C Cell Control is at a) C b) ambient temperature c) either of the above 4. Show your facilitator the control folder printouts showing the following: a. A result that has been removed from the statistical information b. A comment based on a control result c. A set of thumbnail graphs d. The printout showing control runs from one shift. 5. How do we know that a control run is out? Facilitator sign off QC-11 of 12

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51 QC2 ADDITIONAL QC OBJECTIVES Given an operating COULTER LH 500 Workstation, access to HELP, and removable media Download control data for IQAP Demonstrate how to set up and use lab limits Why Is It Important? Using the IQAP program gives your lab a means of peer review with other users of LH 500 systems. Using lab limits allows your lab to more closely monitor control results and take quick action to correct out-of-limits situations. Resources LH 500 Workstation with HELP CD or flash drive IQAP manual QC2-1 of Coulter

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53 INFORMATION / PRACTICE SECTION Downloading for IQAP using Removable Media (CD, Flash Drive, 3.5-inch diskette) Downloading using a CD-R is a two step process. First you must format the CD, then you can use it to perform the download function. You can format and label several CDs so that you always have one available. Be sure to store them carefully. Format a CD to Use for Direct Downloading Step Action Place a blank CD-R in the CD writer drive. (Look at the drives to see which is the writer.) From the workstation desktop, double click on the "Burn CD and DVDs" icon. Select "Make a data CD" and then select the Direct CD option. Follow the path for formatting a CD. You may also refer to the procedure in HELP, Formatting a CD for Direct Archiving. You now have a formatted CD to use directly for downloading purposes. Proceed to IQAP Download Procedure. IQAP Download Procedure Step Action In HELP go to Index 1 Type IQAP Click on the topic Participating Click on 2 Follow the IQAP Download Procedure. Select one control folder for downloading. 3 If there is a problem downloading QC data to your chosen removable media, refer to Troubleshooting in the above HELP topic. 5 Save your download for the skill check. QC2-3 of 10

54 IQAP Submission eiqap Electronic uploading of your IQAP control data files from multiple media types via the internet is now possible using eiqap. You must have an IQAP account set up first. Then you will be asked if you want to use eiqap. To enroll in eiqap, go to www. beckmancoulter.com/qap/index.jsp and select the Hematology tab and follow the instructions for registering and enrolling. If you have an IQAP participant ID, have that handy when you register. If you are not already enrolled in IQAP, it can be done as part of the eiqap enrollment process. Once you have the removable media from the previous exercise, you will log in to the internet site for eiqap and follow screen instructions to upload your data. The advantage to the internet upload is that you will have access to the pool data as soon as the minimum number of pool participants has been reached. You will access and print your own reports via a.pdf format. The internet site provides access to the download for Adobe Acrobat Reader if you do not have it already. Once you enroll all eligible hematology instruments in your laboratory, you access your data through your eiqap account. Also, each institution can have multiple users each with either user or administrator access rights. The first user to enroll the institution will be given administrator access, but other users can also be administrators. Each institution must have at least one administrator. Mail-in IQAP diskette While the transition to e-iqap takes place, the mail-in system will continue. You mail the diskette in a provided envelope and receive an electronic report. Eventually, the mail-in system will be phased out. You will receive notification and a timetable for this as the time approaches. Mail-in should only be a last resort. IQAP prefers to receive the files by , if the customer does not have internet access. Customers who mail in diskettes and do not have an eiqap account will have to indicate whether they want their reports ed or faxed. IQAP no longer mails out printed reports. QC2-4 of 10

55 Other IQAP Considerations COULTER LH 500 TRAINING MODULES One of the features of the control file area is the Shift Clock, which allows data review by shift. If you do have multiple shifts set up, then you need to think about how you want to receive your IQAP reports. If you want to receive three separate reports, one for each shift, do nothing. Your current setup will provide that. If you want to receive one overall report, then you need to change the system setup, before you download: Go to System Setup, Quality Assurance Setup, select the Shifts tab. Deselect the Multiple Shifts checkbox (you will see the shift times disappear). Select OK to save. Now do your IQAP download as described in the section above. After the download, go back to System Setup, Quality Assurance Setup, Shifts tab and select the Multiple Shifts checkbox. The shift times will reappear. Select OK to save. The 5C Cell Control requires monthly downloads as soon after the expiration date as possible or when you have stopped running it. Remember that if there are fewer than 10 runs, you will receive a report, but your data is not included in the pool. This may affect some shifts more than others. In general, Retic-C is not run as often as 5C and may run into the problem of not enough data, especially if submitted by shift. QC2-5 of 10

56 Setting Up Lab Limits Use your instrument Workstation for this part of the module. Step Action At the instrument Workstation, from the Command Center, go to (System Set Up) 1 (Quality Assurance Set Up) 2 Use the line select button to highlight the 5C Normal control folder 3 Select (Setup Lab Limits) button. Enter lab limits as given below. (These limits are used for classroom purposes only. You may wish to establish different limits in your laboratory.) 4 WBC 0.5 MCH 1.2 PLT 25 RBC 0.15 MCHC 1.7 MPV 2.0 HGB 0.6 RDW 1.5 HCT 2.5 MCV Click on (OK) to save. 6 Click on again to close the System Setup [Quality Assurance] window. QC2-6 of 10

57 Step Action Select 7 On the Control Tree Open the 5C folder Open the 5C Normal folder. On the screen to the left of the cumulative results are two buttons: 8 Lets you replace the manufacturer values with the means of your own control runs. Lets you restore the assigned values and expected ranges from the control package insert. Note If the buttons are grayed out, verify that there are at least five runs in the file (this is the minimum required before the buttons become active), if there are at least five runs and the buttons are still inactive then log off the Workstation and log on again. Toggle these buttons now to see how they work. Also notice that the points on the thumbnail (and full-page) graphs move as the changes take place. If you can download control data for IQAP and use lab limits, you are ready to do the Skill Check for this module. A facilitator will check your removable media for the IQAP file. QC2-7 of 10

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59 QC2 ADDITIONAL QC SKILL CHECK Give a facilitator the removable media you used to download IQAP data. Answer the following questions: 1. Does downloading your IQAP data, cause your Workstation data to be deleted? Yes No 2. On your instrument Workstation, show your facilitator the lab limits you set up for 5C Cell Control / Normal. 3. In, go to the 5C Normal control folder, and demonstrate to your facilitator that you can switch between and 4. Why is it important to set up your own lab limits? Facilitator sign off QC2-9 of 10

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61 RC REPLACING COMPONENTS OBJECTIVES Given a COULTER LH 500 or practice station requiring a component replacement, locate the procedure using HELP or use the procedure provided and perform the procedure following all safety precautions. If you complete an exercise at an instrument, ensure the instrument still operates normally. Normal operation is defined as a run of a patient sample without any instrument errors. Why Do I Need To Know This? Knowing how to replace some basic components will minimize instrument down time and require less service calls. Why Can t I Practice Now? Maybe you can! If you are already familiar with the following basic information and tool tips, you may proceed to the practice exercises. Different types of tubing and why they are different. Colors, Lengths and critical lengths Silicon, Polyurethane, Pharmed etc. Pinch valves, mini pinch valves Angar valves Check valves and their sizes, correct orientation Chokes, colors and correct orientation Fittings, metal or plastic Fuses Parts of the Needle Assembly Parts of the Hemoglobin Lamp Assembly The following pages review this basic information. RC-1 of Coulter

62 Resources On the supply table you will find: Display sheet with labeled parts Boxes with different types of tubing, pinch valves, check valves, chokes, fittings, etc. Needle assembly Hgb Lamp Assembly Fuses Screwdriver Pencil You may consult the following topics in Help: Replacing Tubing Checking Pinch Valves Remove the Pinch Valve Tubing and Pinch Valve Replacing Pinch Valves Replacing Check Valves Replacing Fuses Replacing the Needle Assembly (video) Replacing the Hemoglobin Lamp Assembly Note: The sections that follow are not meant to be all-inclusive as to every type or possible application of the parts and pieces covered. The main intent is to provide a familiarity with the different parts and an awareness of the differences that may exist. RC-2 of 16

63 INFORMATION / PRACTICE SECTION TUBING Types of Tubing COULTER LH 500 TRAINING MODULES The general rule of thumb is to replace a piece of tubing with the same type as what you remove. Color-code stripes or printed part numbers identify the proper match. This is important because it ensures that the correct inner diameter of tubing is used. Silicon I-beam Always used in pinch valves through the normally closed part of a pinch valve. I-beam tubing must be threaded properly in the pinch valve tracks. Color-code stripe Blue, Black, Red, No stripe. Round Almost always used in pinch valves through the normally open part of a pinch valve. Color-code stripe Blue, Black, Yellow, Red, Green, Brown. Pharmed A more chemically stable type of tubing (it has an opaque, cream color) I-beam and round versions Color-code stripe Blue, Black, No stripe Polyurethane A harder type of tubing not easily pinched Usually has a part number printed on the tubing If no part number is printed, it will have a color stripe or stripes because it has a particular function, for example: Green/black used in 30 psi/vent lines White used for diluent or cleaner lines Green used in 30 psi lines Special tubing There is some tubing that comes precut, with or without molded ends. It is prepackaged individually and has a specific part number. Tubing Lengths The general rule of thumb is to cut the replacement piece of tubing to the same length as the faulty piece. Tubing designated as critical length is usually marked with a small red plastic ring that slides onto the tubing. There is some critical length tubing that is precut and may or may not have fitted ends attached. This precut tubing is packaged individually and has specific part numbers. RC-3 of 16

64 PINCH VALVES ANGAR VALVES CHECK VALVES Double-action pinch valves are proprietary, patented valves that are designed specifically to work with I-beam tubing. These pinch valves can be removed with a pinch valve wrench (removal tool) to make tubing replacement a little easier or to replace a broken pinch valve. View the HELP topic: Remove Pinch I-Beam Tubing. Use a red transit clip to hold the pinch valve open when replacing I-beam tubing. Remember that I-beam tubing must be threaded properly in the pinch valve tracks. One simple idea to make this easier is to use a two-inch piece of applicator stick, inserted into one end of the I-beam tubing. This makes threading the tubing easier and ensures the I-beam is in the tracks correctly. There are also mini pinch valves (single) and triple (six tube) pinch valves. This type of valve performs a similar function to the pinch valve, but it does not pinch any tubing. The tubing attaches to the outside of the angar valve only. These valves switch one common fitting to one of two other possible fittings internally. Check valves allow liquid or air flow in only one direction. Since they allow flow in one direction only, the orientation of the valve in the line is important. There are three check valve sizes: small, medium and large. Some medium check valves have a black stripe, indicating they have been tested to a certain specification. If a check valve you are replacing has a black stripe, be sure to replace it with a black striped check valve. See Diagram on page RFC-5. When replacing a check valve, remove the tubing one end at a time from the old check valve and reconnect to the new check valve to ensure the direction of flow is correct. RC-4 of 16

65 Check Valves and Chokes (examples) COULTER LH 500 TRAINING MODULES Plastic Choke Check Valves Direction Metal Choke Fitting with choke CHOKES Metal There are five types of chokes, color-coded by sizes, described below. Metal chokes restrict the flow of the air in a pathway. Cylindrical metal chokes An arrow on the cylindrical metal choke represents the direction of flow and should be noted when replacing the choke. These chokes do not come with fittings on them. It is necessary to unscrew the fitting from the end of the old choke and screw it into the new choke. There is an o-ring on the fitting that must be in place to ensure no air leaks at the union point. Color coded Red, Black, Brown, Green, Blue, Gold Metal fittings with a built-in choke Color-coded Blue, Green, Brown, Black, Red Variable Chokes Allow adjustable restriction RC-5 of 16

66 Plastic Plastic chokes restrict the flow of liquid or air in a pathway. There are two styles of plastic chokes. They also have direction indicators. (See figure on page RFC-7) Color-coded Black, Brown, Gray, Yellow, Blue Restrictors FITTINGS Tubing used to provide a certain amount of restriction to the flow of liquid or air. Can be metal or plastic There are several styles including Y, T, Cross-, Feed-through, Union, Reducer. Connects tubing to components or tubing-to-tubing. Carefully push tubing straight onto a fitting. Push tubing onto a fitting so that the end of the tubing is beyond the barb on the fitting. Never use COULTER CLENZ (cleaning agent) or LH Series diluent or CBC Lytic Reagent to moisten the end of a tubing for lubrication. Any residue may dry into a salt or gel and affect instrument operation. You may use Reagent Grade water. You may also carefully stretch the end of a tubing using hemostats, or needlenose pliers or even a toothpick. It depends on the tubing. You must be careful not to create a hole in your new tubing. Fittings (examples) T-fittings Y- fitting RC-6 of 16

67 FUSES The analyzer must have power OFF before attempting to examine or replace a fuse. Always replace a fuse with the same Type and Rating as the suspected defective fuse. (E.g. SLO BLO, 6.25A.) If a fuse fails shortly after you put it in, turn the power OFF to prevent possible damage to the instrument s electrical system. Unplug the power cord from the wall outlet and call your Beckman Coulter representative. Replacement fuses are part of your spare parts kit. Try to save the instrument packing list which has all spare parts listed with their part numbers. LH 500 Specific The fuse panel for the LH 500 is located behind the left side panel. If you have an error message such as VDC (voltage) OUT OF RANGE You may need to replace a fuse. Find the procedure for replacing fuses in HELP. Power down and lift the left panel up and off the left side of the instrument, following the instructions in the HELP procedure. Do not do this now, just locate. Look closely at the fuse panel (use the fuse panel example in class). There are nine fuses labeled F1 through F9 Each shows the type and rating Locate fuse F10 on the back of the analyzer (lower right corner). Replace the left side panel. Power on, following the instructions in the HELP procedure. RC-7 of 16

68 NEEDLE ASSEMBLY INFORMATION This assembly consists of a cartridge holder, a plastic bellows and the needle. At the bottom of the assembly are three lines: Aspiration line Needle vent line (line 1) Waste line (line 3) A new needle assembly includes all three lines already attached. Find and review the procedure for replacing the needle assembly in HELP. If there is nothing wrong with the aspiration line you use presently, just remove the line that comes with the new assembly and transfer the one in use to your new assembly. If you do decide to use the new aspiration line, be sure to run your controls to verify no calibration changes. Always perform the Clean Needle procedure to disinfect the needle before handling the assembly. Be sure the aspiration and vent lines are not twisted. Remember that the assembly is mounted at an angle, so when removing or replacing the assembly, it is important to apply the same angle. Remove the metal needle shield to gain access to the needle assembly. Use the supplied needle safety clip whenever handling the needle assembly. Always Power Off the Main Power switch on the back of the instrument before touching the needle. The two LED indicators on the front of the instrument should be off. Line 3 Line 1 Remove the Safety Clip when finished. Replace the needle safety shield. Power On the instrument. RC-8 of 16

69 HEMOGLOBIN LAMP ASSEMBLY INFORMATION The hemoglobin lamp assembly consists of a lamp and connector mounted in a metal frame. The metal frame has two screw mounts to attach it to the CBC module. Find and review the procedure for replacing the hemoglobin lamp assembly in HELP. Turn OFF the Main Power from the back of the instrument. Disconnect the connector plug. Using a pencil, trace around the assembly at the point it connects to the instrument. Then remove the two screws. Verify the replacement lamp assembly s lamp filament is vertical. (Look through the round opening on the side.) Using the traced outline, attach the new assembly to the instrument with the two screws. (Hint: Use the stick trick to get the screws started. Ask your facilitator) Connect the lamp plug. Turn Main Power ON. Wait 15 minutes and perform the HGB Lamp Adjust procedure. HGB Lamp Adjust Switch to the Instrument computer display by pressing the top button on the switch box. Select Diagnostics Operator Options Hgb Lamp Adjust Press Enter, Enter. The hemoglobin voltage adjusts automatically. RC-9 of 16

70 PRACTICE FOR REPLACING FAULTY COMPONENTS As you progress through this practice session, feel free to ask your facilitator for assistance in accomplishing each task. Repeat any procedure until you feel comfortable doing it. The following five practice sessions (A, B, C, D, E, F) may be done in any order you wish. When you are ready, have a colleague inspect and sign-off your work, then replace the labels indicating the faulty parts. Materials You Will Need Access to the HELP System At the supply table you will find: A practice station with labeled faulty part(s) Spare parts Tools Practice A 1. Locate the leaky tubing in the practice station. 2. Select the correct replacement tubing. 3. Use a red transit clip to hold the pinch valve open. 4. Remove defective tubing and replace with the new tubing. 5. Remove transit clip. Checklist for Practice A The I-beam tubing is the same type and length as the tubing being replaced. The tubing is threaded in the tracks in the pinch valve. Both ends of the tubing are reconnected so they don t leak. Transit clip has been removed. Colleague sign off Continue on next page RC-10 of 16

71 Practice B COULTER LH 500 TRAINING MODULES 1. Locate the faulty check valve in the practice station. 2. Select the correct replacement check valve. 3. Remove defective check valve and replace with the new one. Checklist for Practice B The correct size check valve was used. The check valve is in the original direction. Colleague sign off Continue on next page RC-11 of 16

72 Practice C 1. Locate the clogged needle in the practice station. 2. Follow the procedure for replacing the needle assembly found in HELP and the information found in this module. 3. Disconnect the vent and waste line fittings. 4. Remove the defective needle from the holder. 5. Disconnect the aspiration line from the fitting on the defective needle. 6. Remove new aspiration line from the new assembly and attach the aspiration line from step 5 to the new assembly. Install new needle assembly. 7. Attach vent and waste lines to their correct connectors. Checklist for Practice C The waste and vent lines are reconnected to their correct fittings. The needle assembly is inserted properly in its holder. No lines are pinched or bent. As part of the Skill Check, you will be asked to pull out and then replace a needle assembly at an instrument. No tubings will be disconnected. Colleague sign off Continue on next page RC-12 of 16

73 Practice D COULTER LH 500 TRAINING MODULES 1. Locate the faulty Hemoglobin Lamp Assembly on the practice station. 2. Follow the procedure for replacing the hemoglobin lamp assembly found in HELP and the information found in this module. 3. Unplug the Lamp connector. 4. Using a pencil, trace around the assembly at the point it connects to the instrument. Then remove the two screws that hold it in place. 5. Look at the replacement lamp assembly to verify the lamp filament is vertical. (Look through the round opening on the side.) 6. Plug in the new Lamp connector. 7. Using the traced outline, attach the new assembly to the instrument with the two screws. 8. Verbally describe the next steps for the Hgb Lamp adjust procedure and point to the pathway on an instrument computer display. Checklist for Practice D Traced outline of old assembly before removing to ensure correct alignment. Reattached connector. Correctly described Hgb Lamp Adjust procedure. Colleague sign off Practice E Locate the correct faulty fuse at the practice station based on the following error message: VDC +12 OUT OF RANGE. 2. Follow the procedure for replacing fuses found in HELP and the information found in this module. 3. Remove faulty fuse and replace with correct type and rating. Checklist for Practice E Correct fuse number located. Correct replacement chosen. Colleague sign off If you feel confident with these skills and all sign-offs are complete, you are ready to proceed to the Skill Check. RC-13 of 16

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75 RC REPLACING COMPONENTS SKILL CHECK Using a COULTER LH 500 or practice station in the lab, locate and perform the following replacement procedures: Replace the leaky tubing in a single pinch valve Replace a check valve Replace a Needle Assembly Replace a Hemoglobin Lamp Assembly Replace a fuse All necessary tools and materials can be found on the supply table. The replacements may be done in any order. When finished, ask a facilitator to inspect your work for sign-off. Inspection Checklists Checklist for Leaky Tubing Replacement The I-beam tubing is the same type and length as the tubing being replaced. The tubing is threaded properly in the pinch valve. Both ends of the tubing are properly reconnected. Transit clip has been removed. Checklist for Check Valve Replacement The correct size check valve was used. The check valve is in the proper (original) orientation. RC-15 of 16

76 Checklist for Needle Assembly Replacement Select correct needle type. The aspiration line is reconnected to the front blood detector. The waste and vent lines are reconnected to their correct fittings. The needle assembly is inserted properly in its holder. No lines are pinched or bent. Direct observation at their instrument of student disconnecting needle shield, pulling needle assembly out of the holder and then replacing properly. Student then replaces needle shield. Checklist for Hemoglobin Lamp Assembly Replacement Traced outline of old assembly before removing to ensure correct alignment. Reattached connector Described Hgb Lamp Adjust procedure correctly. Checklist for Fuse Replacement Correct fuse number located. Correct fuse replacement chosen. Facilitator sign off RC-16 of 16

77 RPS RUNNING PATIENT SAMPLES OBJECTIVES Given an operating COULTER LH 500, bloods collected in EDTA, other supplies, and access to HELP State the maximum aspiration volume needed for automatic and manual modes of operation State the maximum stability of CBC/Diff and Retic samples stored at room temperature and stored refrigerated Identify the two types of cassettes that can be used on the LH 500 and the proper tubes to use in each Process bar coded samples in the automatic mode of operation and print results automatically Process bar coded and non-bar coded samples in the manual mode of operation and print results automatically Process a STAT sample Process samples in the Pre-dilute mode Identify the populations on the two-dimensional dataplot for Diff and Retic Find samples in the Database using Navigation buttons (Previous) Why Is It Important? Running samples is a daily task that occupies much of your time in the lab. Learning how to process samples efficiently on your LH 500 will save time. Resources To complete this module you will need LH 500 LH 500 Workstation with HELP Blood samples collected in EDTA anticoagulant Bar code labels (located in trays on the Skill Check cart) Cassettes Pipettors, tips and clean tubes (for Pre-dilute) LH Series Diluent Microtainers (for elective exercise) RPS-1 of Coulter

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79 INFORMATION / PRACTICE SECTION Sample Storage and Stability 1. At a Workstation, from (HELP), go to Contents LH 500 Analyzer Operating Operating-BASIC 2. Display and print the following topics Collecting Specimens Storing Specimens. 3. Close HELP. 4. Answer the following questions: What is the minimum amount of properly collected venous sample required in a sample tube to be able to run in the Automatic mode? What is the maximum volume of blood aspirated in the Automatic mode? In the Manual mode? State the maximum stability of a CBC/Diff sample stored at Room temperature refrigerated State the maximum stability of a Retic sample stored at Room temperature refrigerated 5. Have a facilitator review the different cassette choices available for use on the LH 500. Automatic Mode Before you begin, make sure you have the supplies you need: Blood samples collected in EDTA Bar code labels Sample cassettes Be sure to wear your labcoat and gloves, during this procedure. Instrument/Workstation Overview 1. Obtain bar code labels for CD, C and R Run Types from the specified baskets and place them on the sample tubes at your unit. 2. Click on (Run Configuration) from the Command Center. Make sure that Automatic Output, Print (tab) is set to All Samples 3. At the Command Center, select AUTO ANALYSIS as the Process Type, CD as the Run Type and AUTO as the Asp Mode. Select Start. RPS-3 of 10

80 Running Samples 1. Place CD sample tubes into a sample cassette, making sure the bar code labels are facing up. 2. Place the cassette into the right-hand loading bay. A sensor detects the presence of the cassette and starts the run automatically. If nothing happens look at the Workstation Instrument Status box to see if the instrument started. If not, press the Start button. 3. Look at your printouts. Compare to the Patient Tests [Results & Graphics] window. 4. Click on the Diff Data tab. 5. Pass the cursor over the populations on the the two-dimensional Dataplot to learn their identification through the ToolTips. 6. Double-click on the two-dimensional Dataplot to enlarge it. 7. Close the Dataplot by selecting [Close] 8. Repeat exercise using the C labels and make appropriate changes tocommand Center. Manual Mode Using the hand-held scanner to identify samples Be sure to wear your full-face protection, in addition to your labcoat and gloves, during this procedure. 1. Place a C or CD bar code label on a sample. Change Run Type appropriately and change Asp Type to Manual. Select Start 2. Cursor automatically goes to the Barcode field. 3. Hold the scanner over the bar code label. Press and hold the trigger until the scanner beeps. The barcode number appears in the Barcode field. 4. Make sure the sample is well mixed. Remove the stopper from the tube. 5. Immerse the manual mode probe in the sample. Press and release the aspiration bar to activate manual mode aspiration. When you hear the beep from the monitor, remove the tube and re-cap it. The probe wash rinses and dries the probe automatically. NOTE: If you have a non-bar coded sample, you can manually type the accession or identification number into the Barcode field on the Command Center by using the keyboard. Press <ENTER> or <TAB>.TRY IT! RPS-4 of 10

81 Running a STAT COULTER LH 500 TRAINING MODULES 1. Obtain four CD bar code labels. Set up Command Center appropriately for Auto aspiration mode. 2. Put three labeled tubes into a cassette and place in loading bay. 3. After the first tube aspirates, select Stop from the Command Center. 4. When the cycle completes, select Manual Asp Mode. Select Start. 5. Run the Stat in manual mode. When the cycle completes, select stop. 6. Resume cycling the cassette by selecting the Auto Asp Mode, then Start. Using the Predilute Mode The Predilute mode on the LH 500 may be used to run specimens that are over the reportable range or to run a citrated tube when you have clumped platelets. When chosen, it runs in the CBC test mode only. You may enter dilution factors from 1.1 to 5.0. The sample results are automatically multiplied by the dilution factor entered. The minimum amount of blood with which to make a dilution is 50 µl. Remember that Manual mode requires 125 µl of sample for aspiration. After running a dilution, the Analyzer automatically disables predilute. Use a sample for which you already have results. 1. Make a dilution manually using LH SERIES DILUENT provided. Using a pipettor: Dispense 100 µl blood into a clean tube. Dispense 100 µl LH Series Diluent into the same tube, and mix well. 2. On the Workstation, Command Center: Select Stop Predilute (CBC) is enabled Set the Factor to 2.0 in the pop-up box that appears for the dilution just prepared. Select Start Note: There is no need to change Run Type or Asp Mode. This allows you to continue using your previous settings once the Predilute cycle completes. 3. Identify your sample to the LH 500 using the keyboard to type an ID. Press Enter or Tab. 4. Run your sample in the Manual mode. (Predilute is only active in the manual mode.) 5. Print your results. 6. Check these results with the results of the same sample run undiluted. 7. Save printouts of the undiluted and diluted results for the Skill Check. REMEMBER: Setting a dilution factor is valid only for one sample. It automatically reverts back to 1.0, once the cycle completes. RPS-5 of 10

82 Using Micro-collection Tubes (Elective Exercise) Get micro-collection tubes, EDTA blood tubes, and transfer pipettes from the supply in the lab. 1. Look at a demo micro-collection tube for the fill level required for manual mode aspiration. 2. Using a transfer pipette, fill two or three micro-collection tubes with blood from one of the sample tubes you already have. 3. Get bar code labels for CD and C Run Types. 4. In Run Configuration, set Print to All Samples. 5. Use the hand-held bar code scanner to enter the bar code ID# at the Workstation. Refer to the Manual Mode procedure on page RPS Process sample in the manual mode. 7. For additional samples, repeat steps Save printouts for the Skill Check. Running Samples in the Retic Mode (Elective Exercise) The Reticulocyte methodology on the LH 500 is semi-automated. It requires the use of the Retic Prep Kit (PN ) which contains two reagents, A and B. Reagent A modified New Methylene Blue stain provided in a dropper bottle Reagent B a clearing solution of very dilute sulfuric acid The preparation steps must be followed exactly for accurate results: 1. Use one or two samples. 2. Label two 12 x 75 ml glass vials for each sample. 3. Pipet 50 µl of well mixed sample into its labeled vial. 4. Add four drops of Reagent A immediately and shake vial to mix blood and stain. Allow to incubate at room temperature for a minimum of 5 minutes and a maximum of 60 minutes. 5. Prepare instrument to run the Retic mode: Select. Choose Process Type AUTO ANALYSIS and Run Type R. Be prepared to type or scan the ID. Select 6. Gently remix a blood/stain vial. Pipette 2 µl of the mixture into the bottom of the clean, labeled 12 x 75 ml glass vial for that control. 7. Immediately dispense 2 ml of Reagent B into the vial, holding the vial at an angle while doing so. This method provides mixing of the sample with the clearing solution. Do not shake to mix. 8. Wait 30 seconds and then analyze the retic preparation you just made. Type in the ID and press Enter or Tab. Immerse aspirator tip into the retic preparation Press and release the sample bar (it will aspirate the entire contents) Remove the tube when you hear the beep 9. Review results. RPS-6 of 10

83 Using Navigation Buttons 1. At the Workstation, on the Patient Tests [Results & Graphics] window, use the Navigation drop-down box to select the Completed folder in which you want to look at samples. Example: CBC/Diff. It acts as a sort, using the Navigation buttons. 2. Use Navigation buttons (located on the Specific Toolbar) to find samples that you have run previously today. How to Rerun a Sample Using the ToDo List (Elective Exercise) When you are bidirectionally interfaced, sample requests are automatically downloaded to the LH 500 ToDo List by your Host/LIS system. When you run the samples, they are removed from the ToDo List. If you need to rerun a sample there are two scenarios: 1. Just rerun it. The Workstation gives you a No Match status along with the Sample ID (from the bar code label) and the results. The instrument uses the Run Type mode selected at the Command Center. There will not be any demographics associated with the rerun. You can always edit the demographics to the run later. 2. Manually add it back to the ToDo List. This option eliminates the No Match status and runs the sample in the preassigned Test mode. If you want demographics they have to be entered at the time you add it to the ToDo list. RPS-7 of 10

84 Adding a Sample Request to the ToDo List 1. Select (Add Sample Requests) from the common toolbar of the patient results area. 2. Select a Test Mode check box. 3. Click once in the Sample ID area to place your cursor, then scan the tube bar code label. Leave the Cassette/Position field blank. 4. Use the <Tab> key to move the cursor to the Patient ID field. 5. Open the drop-down box to select the Patient ID. Since this is a previously run sample, it should be there. The other demographics associated with the sample automatically populate the fields. 6. Add any other information you may need and then select at the bottom of the screen to add it to the ToDo list. 7. Select (exit). If you wish to confirm the new entry, go to (Database/ToDo View) and then select the test mode folder you assigned to the repeat. 8. Place the repeat tube into any cassette and run it. Proceed to the Skill Check for this module. Bring required printouts to a facilitator as part of the Skill Check. RPS-8 of 10

85 RPS RUNNING PATIENT SAMPLES SKILL CHECK Have your facilitator check your answers to the questions on page RPS-3 of the module. Check the tube types and sizes that may be used in each type of sample cassette: 13 mm cassette (gray) 16 mm cassette (black) Controls S-CAL Hemogard 2 & 3 ml pediatric 5 ml rubber stopper 7 ml rubber stopper Show the printouts from this module to your facilitator. At an LH 500 Workstation, use the Navigation buttons to find a twodimensional diff Dataplot on a sample that you ran for this module. Using HELP, if necessary, show your facilitator that you can identify Diff populations and enlarge the Dataplot. Which of the three Run Types is used when you run a Prediluted sample? If you were running a citrated tube because of clumped platelets, what would you type in as a dilution factor? Facilitator sign off RPS-9 of 10

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87 RSR REAGENT SET UP AND REPLACEMENT OBJECTIVES Given a COULTER LH 500 Workstation, access to HELP, and Handheld Barcode Scanner. Update the Reagent Setup screen in the LH Workstation (if necessary.) Recognize instrument messages used for reagents. Properly change a reagent container. Properly replace a waste container. Why Is It Important? An up-to-date reagent setup screen shows that your laboratory is aware of good Quality Assurance procedures. Your LH Workstation makes the task of entering new reagents easy. Resources To complete this module you will need: An Individual LH 500 Workstation with HELP and a Handheld Barcode Scanner New reagents RSR-1 of Coulter

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89 INFORMATION / PRACTICE SECTION The Reagent Data Entry screen shows the current reagent lot numbers in use on your instrument. In addition, this screen shows the unopened shelflife expiration date (the date on the box) and the open container expiration date. When you make an entry in the Reagent Setup screen, there is also an entry made in the History Log Reagent tab that includes the date and time of the reagent change, along with the lot number and expiration date. Reagent Data Entry Screen The Shelf Life is the date to which an unopened container may be used. These are the default open-expiration dates for Beckman Coulter reagents. The open expiration is the date to which an opened container may be used, based on the Date Opened. RSR-3 of 10

90 Reagents Used on the LH 500 Series Diluent The isotonic diluting fluid is LH Series Diluent (PN ) CBC Lyse The CBC lysing fluid is LYSE S III diff (PN , 1L; , 5L) Diff Pack The differential reagents are LH Series PAK (PN ) Cleaner Pack The cleaning agent is COULTER CLENZ (PN ) Retic Prep Kit Reagent kit to prepare a sample for reticulocyte analysis is Retic Prep Kit (PN ) Setting Up a New Reagent Step Action In HELP go to SEARCH Type reagent 1 Click on List Topics Select Changing Reagent Information Click on Read the procedure Close HELP On the instrument Workstation from the Command Center go to (System Set Up) 2 (Quality Assurance Set Up) The tab is the first tab on the QA Set Up window. 3 Select the button to setup new or modify existing reagents. The Reagent Setup window appears. 4 Enter the reagent information by using the handheld barcode scanner or manual entry of the barcode. Note Press tab to move between fields for manual entry. RSR-4 of 10

91 5 Select (OK) to save and exit the Reagent Setup window. 6 Select in the Reagent Data Entry window to save the changes. NOTE: When the audible reagent alarm signals that a reagent container is out, you must replace the reagent container immediately. The Workstation automatically displays a red message box. When you acknowledge this message by selecting OK, It closes the message and the traffic light returns to green. The reagent low message is also posted to the History Logs, Instrument tab. RSR-5 of 10

92 The Workstation displays these messages for the reagent that is out: CLENZ OUT DILUENT OUT LYSE OUT PAK OUT WASTE FULL (Only applies if you collect waste into a waste container.) Steps to Change a Reagent Container Turn the compressor Off Switch to the Instrument Computer display by pressing the top button (Position 1) on the switch box. From the Main Menu select Diagnostics Operator Options Fluidic Tests Compressor On/Off Press Enter, then Spacebar, then Enter again. Open the new reagent container. Remove the pickup tube assembly from the old container and transfer it directly to the new container without touching anything. Tighten cap. Note: If the lower part of the assembly touches you or anything outside the container, flood that lower part with distilled water then wipe it with lint-free tissue. Prime the Reagent From the Main Menu select Diluter Functions Prime Reagents Press Enter, select the reagent that was changed, press Enter Switch back to the Workstation display by pressing the bottom button on the switch box. Verify the Process and Run Type selections and press Start to continue running samples. RSR-6 of 10

93 Steps to Replace a Waste Container If you collect instrument waste into a container, then you will need to follow these steps when you get a Waste Full message. Put the instrument into Standby Move the Standby/Reset switch to Standby (position O). Turn OFF the Main Power using the rocker switch on the back of the instrument. Replace the Waste Container Have an empty container ready. Using biohazard precautions, remove the waste assembly from the full container and transfer directly to the empty container. Dispose of the biohazardous waste according to your laboratory s protocol. Turn ON the Main Power Put the instrument into Ready Move the Standby/Reset switch back to Ready (position I). Resume operation. Proceed to the Skill Check. Be ready to show your facilitator the reagent log that you set up on an Individual Workstation. Remember that during class you are responsible for changing reagents and up-dating the reagent log on the LH 500 Workstation you are assigned to. RSR-7 of 10

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95 RSR REAGENT SET UP AND REPLACEMENT SKILL CHECK 1. Demonstrate the steps to update the reagent information for a new container of reagent to your facilitator. 2. If you replace a reagent container, how do you Prime the reagent? 3. When you replace a reagent container you must turn the Off. 4. When you replace a waste container, you must turn the Off. Facilitator sign off RSR-9 of 10

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97 SC1 SETTING UP CONTROLS I OBJECTIVES Given a COULTER LH 500, access to HELP, control package inserts and diskettes Access the screen for setting up new control folders on the LH 500 Workstation Set up COULTER LATRON Control folder using data from the package insert Set up COULTER 5C Cell Control and COULTER Retic-C Cell Control folders using either the removable media, optional 2D Barcode Scanner or the Control diskette (to be phased out) Explain how a lab might use Auto Transmit and Auto Stop features Why Is It Important? Proper and timely set up of control folders is the basis of a good lab QC program. Resources To complete this module you will need An Individual LH Workstation with HELP and access to the internet LATRON package insert from the facilitator Removable Media (CD, Flash drive or 3.5 diskette) SC1-1 of 8 Coulter

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99 INFORMATION / PRACTICE SECTION COULTER LH 500 TRAINING MODULES Step 1 2 Action Get removable media for downloading control assigned values and expected ranges from the Internet and a LATRON Control package insert. At your INDIVIDUAL Workstation, open the internet and access the following pathway to download control assay values. support>qa & Service Programs>Go to eiqap>hematology Assays. 3 Follow the on-line instructions for downloading. 4 Once the control assay values are downloaded to the removable media, follow the next steps. From the Command Center go to [System Set Up] 5 [Quality Assurance Set Up] (tab) Line Select Set up Lab Limits Set up New Control Folder Archive Edit values Transmit Delete selected files SC1-3 of 8

100 NOTE: The Enable 2-Page Run List Printouts option allows for printouts for the CD control in a larger font. When this is selected, the printout is two pages wide vs. one page wide. In either case, there may be multiple pages depending on the number of runs in the control folder. 6 Select [New Control Folder] 7 Choose the Type, Source, and Level of control folder you want to set up. BCI 5C All Levels BCI Retic-C All Levels BCI LATRON 8 Select [Set Up New Lot] 9 If you are setting up either 5C Cell Control or Retic-C Cell Control, follow directions on the pop-up window. You may select to have the LH 500 Analyzer AutoStop when a control is out of limits. If you select AutoStop, go to [Run Configuration] and also select AutoStop Criteria/ Controls. You may select to AutoTransmit control results to a Laboratory Information System if the LIS has a QC package. 10 Select [OK] 11 Warning When you are setting up LATRON, after selecting the button, you enter information from the package insert starting with Lot # and expiration date. Use the Tab key to move from field to field. If you press the Enter key, the screen will close. To continue, you need to select the Latron file from the control list by using the line select button and then select the Edit button. You do not have to enter the (+/-) or ( ) signs when typing numbers. 12 Select [OK]. 13 Select [Close] to close the Setup New Control Folder window. 14 Select [OK] to close the System Setup / Quality Assurance window. SC1-4 of 8

101 ALTERNATE METHOD (MID 2010) COULTER LH 500 TRAINING MODULES In 2010 an additional method for setting up controls will be available using the optional 2D- Barcode Scanner. Assay sheets for 5C and Retic C controls will contain 2D barcodes which when scanned will set up all three levels of controls. Have you set up all required control folders? You should have four or seven control folders. If YES, proceed to the Skill Check. Show a facilitator the control folders you have set up on your Individual LH Workstation. In addition, be able to tell a facilitator how the Auto Transmit and Auto Stop features work. If NO, go back and set up the missing control folders. SC1-5 of 8

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103 SC1 SETTING UP CONTROLS I SKILL CHECK 1. Show a facilitator the four or seven control folders you set up on your Individual Workstation. 2. How might a lab use the Auto Transmit feature? 3. How might a lab use the Auto Stop feature? 4. The facilitator will ask you to delete all the folders after they have been checked. Locate the delete button on the screen, but do not use it until the facilitator asks you. You may use Help to assist you if you don t see it. Facilitator sign off SC1-7 of 8

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105 SD SHUTDOWN OBJECTIVES Given an operating COULTER LH 500 Perform the Clean procedure using COULTER CLENZ cleaning agent State what cycle is performed at the end of the 30 minute Clean time State the difference between the Clean procedure and the Shutdown procedure State the minimum time for the LH 500 to be in Shutdown State the maximum time for the LH 500 to be in Shutdown State when it is necessary to perform Extended and Prolonged Shutdown Why Is It Important? To ensure optimal cleaning of the instrument, the Clean procedure or the Shutdown procedure must be followed daily. Resources To complete this module you will need LH 500 SD-1 of Coulter

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107 INFORMATION / PRACTICE SECTION COULTER LH 500 TRAINING MODULES The LH 500 must be shutdown in a cleaner (CLENZ) cycle for a minimum of 30 minutes once every 24 hours of use. The shutdown process removes reagents from the instrument and replaces it with cleaner. The Clenz reagent is an enzymatic cleaner that rids tubing of debris and prevents protein buildup on apertures, in the BSV (blood sampling valve) and in the flowcell. The LH 500 gives you two ways to accomplish this: The Clean cycle that switches the instrument to cleaning reagent, waits 30 minutes and then automatically performs the Startup cycle. OR The Shutdown cycle that switches the instrument to cleaning reagent. The operator must then request a startup after 30 minutes or more. The Workstation posts an entry to the Daily Checks History Log with the date and time that the Shutdown request occurred. Clean Cycle Please perform this procedure every afternoon in the classroom at about 4: From the Command Center select Stop to put Instrument Status to SELECT FUNCTION. 2. Switch to the Instrument Computer display by pressing the top button at the switch box. 3. From the Main Menu, select Diluter Functions Clean. 4. Press Enter, Enter. The screen displays the message Clean Cycle in progress. This cycle takes approximately 35 minutes. Please wait A timer displays in the upper right. Instrument Status display is SHUTDOWN and then SYS NOT READY. It takes about two minutes to change to the cleaner. Then the system waits for 30 minutes when it automatically begins the startup cycle. This takes about five minutes. Instrument status now displays STARTUP. 5. Switch to the Workstation display by pressing the bottom button at the switch box. You may do this at any time after beginning the Clean cycle. 6. Verify your Daily checks information from the startup. If you need to interrupt the Clean cycle you have two choices F4: Abort the cycle F5: Run Startup now It is preferable to choose F5 if you can not wait the entire 30 minutes. (e.g. a stat request) Note: Clean Cycle timing is NOT programmable or changeable, other than interrupting if necessary, as indicated above. SD-3 of 6

108 Shutdown 1. From the Command Center select Stop to put Instrument Status to SELECT FUNCTION. 2. Select Shutdown from the Process Type box and then select Start to begin the shutdown cycle. Allow the instrument to remain in shutdown for a minimum of 30 minutes. Leave the power ON. 3. Select Startup from the Process Type box and then select Start to begin the startup cycle. 4. Verify your Daily checks information from the startup. Important Notes Once the instrument is in Shutdown, the only allowable Process type is Startup. All other choices are grayed out. Although the minimum time in cleaner is 30 minutes, you may leave the instrument in cleaner for up to 48 hours. If the instrument is left in Shutdown for 23 hours or more with the power ON, the system will automatically perform an Autopurge cycle which: Brings up the compressor Purges the flow cell and sample line with diluent Turns OFF the compressor. This process repeats every 24 hours until you request a startup. If the system will be idle for more than 48 hours (e.g. from Friday afternoon to Monday morning) with the power OFF perform the Prolonged Shutdown procedure. If the system will be idle for more than seven days, perform the Extended Shutdown procedure If the system will be idle for more than 30 days, contact your Beckman Coulter representative for specific instructions. The following two procedures are for your information only. Do not perform them in the classroom at this time, but be familiar with them if they apply to your laboratory. Open Help, find the Prolonged Shutdown and Extended Shutdown procedures. Print and read. Proceed to the Skill Check for this module SD-4 of 6

109 SD SHUTDOWN SKILL CHECK 1. What is the minimum time for the LH 500 Analyzer to be in shutdown every 24 hours? 2. At the end of the Shutdown time in the Clean cycle, what cycle does the LH 500 Analyzer perform automatically? 3. What steps do you take if the Clean cycle is underway, and you get a STAT? 4. What is the difference between the Shutdown cycle and the Clean cycle? 5. How long can you leave the instrument in Shutdown? 6. If the instrument is left in Shutdown to 23 hours, what cycle occurs automatically? 7. What are the conditions to perform the Prolonged Shutdown procedure? 8. What are the conditions to perform the Extended Shutdown procedure? Facilitator sign off SD-5 of 6

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111 SF SAMPLE FLOW OBJECTIVES Given an operating COULTER LH 500, Summary Sheet diagrams, blood sample, and access to HELP Locate and name major Diluter components, using no references List the constituents of WBC and RBC/Plt, dilutions, using references State the four constants and one variable used for the COULTER Principle, using references State the importance of sweep flow and mixing bubbles, using references State the purpose of cross-rinse at the end of an instrument cycle, using references Why Is It Important? Knowing component names and locations as well as their functions is important when you are troubleshooting. Also knowing what happens when a component does not work helps you to troubleshoot. Using the HELP system as a primary resource can help you solve problems. Resources LH 500 Analyzer LH 500 Workstation with HELP Reference Information / Operation Principles Identifying System Components Summary Sheets: Diluter diagrams A thru J (laminated sheets) Sample to run, and magnet from facilitator Component table with instrument "pieces and parts" Self-Check As you complete this module, you will find Self Checks along the way. There is an Answer Key on page SF-27. SF-1 of Coulter

112 SET UP Complete this module with a partner, if possible. Review the module thoroughly before you begin. Be sure to wear full-face protection (either a full-face shield OR protective eyewear with a facemask) along with lab coat and gloves. 1. At the Command Center, select Run Type C and verify the number of aspirations per tube is At the Command Center, go to Set Automatic Output / Print to DISABLED 3. Open the front door panel by pulling towards you from the right side. 4. Remove the needle shield. Loosen the screws on either side of the needle shield and then remove. Ask facilitator for help if needed. 5. Open the right side door pulling from the left. 6. For cycling the LH 500 Analyzer, use the tube provided by a facilitator. 7. Ask facilitator to apply the magnet. When operating the LH 500 Analyzer in your laboratory you must heed the warning shown below. For training purposes, your facilitator will place a magnet on your instrument to defeat the door interlock. You may then run the instrument with the front door open. WARNING Keep the front door of the LH 500 Analyzer closed while you use the Automatic aspiration mode. The system will stop if you remove the needle shield. SF-2 of 30 PN CA September 2009)

113 INFORMATION / PRACTICE SECTION Module and Component Identification COULTER LH 500 TRAINING MODULES The LH 500 consists of ten modules which contain the components to analyze samples. The ten modules are: Analyzer Module Contains the various circuit boards needed to operate the instrument. Electronic Power Supply Supplies the necessary electrical power for all instrument operations. Pneumatic Power Supply Houses the compressor that provides the vacuums and pressures used in the instrument. CBC Module Contains all the components necessary for CBC Analysis. BSV (Blood Sampling Valve) Module Contains the components associated with aspirating blood samples. Pump Module Contains the two Erythrolyse II (Diff Lytic) reagent pumps needed for differential analysis. Autoloader Module Contains the robotic mechanisms to move the cassettes from the loading bay to the piercing station and then to the unloading bay. Mixing Module Contains the components associated with WBC differential analysis Flow Cell Module Contains the components used to perform VCS analysis. Main Diluter Module Contains various components relating to sample dilution and cycling as well as vacuum and pressure regulators. SF-3 of 30

114 Modules Located in the Front (Diagram A) Analyzer Module CBC Module* Electronic Power Supply BSV Module * The appearance of some parts may be different on your instrument. Autoloader Module Pump Module Pull open the front door of the instrument. It opens from the right. Analyzer Module Electronic Power Supply Autoloader Module with Needle Shield removed CBC Module BSV Module Pump Module C aptive S crew s Needle Shield SF-4 of 30 PN CA September 2009)

115 Modules Located on the Back (Diagram B) Fans Reagent manifold and connectors Power ON/OFF rocker switch Plugs for computer Pneumatic Power Supply Go around to the back of the instrument. Pneumatic Power Supply Main Power ON/OFF rocker switch Reagent manifold and connectors SF-5 of 30

116 Modules Located on the Right Side (Diagram C) Main Diluter Module Mixing Module Flow Cell Module Pull open the right side door. It opens from the left. Main Diluter Module Mixing Module Flow Cell Module SF-6 of 30 PN CA September 2009)

117 Analyzer Module (Diagram A) Analyzer Module The Analyzer Module contains the various circuit boards needed to operate the instrument. The various boards are responsible for controlling The timing and sequencing of the operating cycles Receiving pulses and raw data from both the CBC and Flow Cell Modules Counting measuring and computing CBC parameter results Sending final CBC parameter results and VCS data to the Workstation. Electronic Power Supply (Diagram A) Electronic Power Supply The Electronic Power Supply provides both AC Power and DC Power to various components to provide for instrument operation. Pneumatic Power Supply (Diagram B) Pneumatic Power Supply The compressor/vacuum pump produces one service adjustable level of air pressure (60 psi) and one nonadjustable level of vacuum (>22 Hg). SF-7 of 30

118 Autoloader Module (Diagram D) Locate the following components using your laminated sheets as a reference. Rotate the rocker bed forward (push down on the front). Load Stack Empty sensor Load Gate Position detect sensors Load Elevator Tube Available sensor Tube Forward sensor Needle Assembly Unload Elevator Unload Gate Unload Stack Full sensor CBC Module (Diagram E) Locate the following components using your laminated sheets as a reference: RBC vacuum isolator chamber (The appearance of some parts may be different on your instrument.) WBC vacuum isolator chamber CBC Lytic reagent pumps (two pumps, 1.0 ml total) Waste chamber RBC aperture bath WBC aperture bath Overflow chamber Hemoglobin Lamp Hemoglobin pre-amp BSV Module (Diagram F) Locate the following components using your laminated sheets as a reference: Bubble-maker pinch valves Sheath Tank Air Pump Automatic mode aspiration pump Manual mode aspiration pump Front blood detector Rear blood detector Blood Sampling Valve (BSV) Pump Module (Diagram G) Locate the two Erythrolyse II reagent pumps using your laminated sheets as a reference. SF-8 of 30 PN CA September 2009)

119 Mixing Module (Diagram H) Locate the following components using your laminated sheets as a reference: Mixing chamber StabilLyse pump Flow Cell Module (Diagram I) Locate the position of the Flow Cell Module using your laminated sheets as a reference. Also look at the following parts on the component table: Flow cell Laser Light scatter detector Connectors for DC and RF signals VCS electronics Laser on indicator Main Diluter Module (Diagram J) Locate the following components using your laminated sheets as a reference: WBC diluent dispense pump RBC diluent dispense pump Vacuum trap Sample pressure regulator Sheath pressure regulator Backwash pump 30 psi regulator Low vacuum regulator Solenoids SF-9 of 30

120 AUTOMATIC MODE Transport Unload stack full sensor Tube Ram Bar Code Load stack empty sensor Unload gate Load gate Position detect sensors Position detect sensors Unload elevator Load elevator Tube forward sensor Observations Needle assembly Tube available sensor At the Autoloader Module, look at the sample cassette loading bay to the right. Find the Load stack empty sensor. This is a cone-shaped, spring-loaded sensor that detects when a cassette is placed on the Load gate. This activates the Load elevator to move up to the waiting cassette. Find the load gates. The load gates are located on each side of the cassette. The cassette rests on the load gate before the cassettes move to the rocker bed. When the load elevator moves up to pick up the waiting cassette, it pushes the load gate in. After the cassette moves down, the load gate springs out. SF-10 of 30 PN CA September 2009)

121 Look at the rocker bed. Find the load elevator. The load elevator moves through an opening in the rocker bed to pick up a sample cassette from the loading area. Find the tube available sensor. It is a cone-shaped, spring-loaded sensor located near the center of the rocker bed. When the sensor detects a tube in a cassette, the cassette locks in place. Find the tube ram and the tube forward sensor. The tube ram is a silver cylinder located underneath the bar code reader. Once the cassette locks in place, the rocker bed tilts to the forward position and the tube ram pushes the tube down. When the tube forward sensor activates, the needle is pushed up piercing the stopper. Find the position detect sensors behind the unload elevator. When the sample cassette activates these switches, it signals the unload elevator to raise the cassette to the unload gates. Find the unload stack full sensor. It is a cone-shaped, spring-loaded sensor located on the right side of the unload bay. When there are five cassettes in the unload bay, the unload stack full sensor generates and error message reminding the operator to remove cassettes from the unload bay. Activity 1. Put the sample tube provided into a cassette and place in the right loading bay. 2. Observe cassette transport. Self Check 1. How does the instrument know that a cassette is waiting to process? 2. What component causes the cassette to lock into position? 3. What is the function of the tube ram? When you finish the Self Check for each section, check your answers with the Answer key on page SF- 27. SF-11 of 30

122 Automatic Mode Aspiration Front blood detector Automatic mode aspiration pump Blood detectors Bubble-maker pinch valves (2) Aspiration line Observations In the automatic mode, the automatic mode aspiration pump draws a maximum of 185 µl of sample through the needle, front blood detector, Blood Sampling Valve, and the rear blood detector. Locate the automatic mode aspiration pump (PM 4). Locate the BSV Locate the Front and Rear Blood Detectors SF-12 of 30 PN CA September 2009)

123 Locate the needle assembly. The needle is contained within a bellows and has two openings near the piercing tip: One for aspiration on the bottom side One for venting on the top side. Piercing needle Vent Sample aspiration (and backwash) Front support Vent line (and rinse) Bellows Aspiration line (and backwash) Waste line The venting pathway in the needle equilibrates the tube contents to atmospheric pressure. If this is not done and there is residual vacuum in the sample tube, it could cause aspiration errors. This pathway is then rinsed with diluent and dried by high vacuum, preventing carryover to the next sample. The aspiration port opens a pathway so that vacuum applied by the automatic mode aspiration pump can pull a sample of whole blood from the specimen tube through the BSV and blood detectors. Examine the needle more closely at the component table in the lab. The blood travels from the base of the needle through a small black box (front blood detector) to the Blood Sampling Valve (BSV) and then to the rear blood detector. Locate this pathway on your instrument. SF-13 of 30

124 Aspiration 28 µl diff loop 1.6 µl RBC 28 µl WBC loop From the BSV the blood continues to the rear blood detector. When the center section of the BSV rotates the blood segments in the BSV into three portions (one for RBC/Plt, one for WBC/Hgb and the third for the Diff). Summarized Steps of Automatic Mode Aspiration The instrument puts the tube into the piercing position. The bubble-maker pinch valves activate and pinch the tubing of the aspiration pathway, thus expelling a drop of diluent from the tip of the needle. The bubble-maker pinch valves deactivate and an air bubble forms in the tip of the needle. The needle pierces the tube stopper and the automatic mode aspiration pump aspirates 185 µl of sample from the tube. The diluent that was in the line, followed by the bubble and the blood segment is pulled by a vacuum created by the aspiration pump. A diluent-bubble-blood interface can be observed. The needle retracts from the tube. Vacuum pulls the blood segment through the front blood detector, the BSV and the rear blood detector. As the vacuum equalizes the diluent-bubble blood moves through and stops outside of the rear blood detector. The blood segment is now in the correct position for the blood detectors to assess the quality of the aspiration and determine if the aspirated blood is acceptable. Activity 1. Put the sample tube provided into a cassette and place in the right loading bay. 2. Observe sample aspiration. Look for the diluent-bubble-blood interface. Self Check 1. State one problem that could result from a plugged needle vent. 2. What is the aspiration volume for the automatic mode? SF-14 of 30 PN CA September 2009)

125 Sample Delivery During aspiration, the aperture baths, containing diluent rinse from the last cycle, drain to the waste chamber. This prepares the baths for the new sample dilutions. Draining the Baths FF3 FF5 The appearance of some parts may be different on your instrument. Check Valves Waste Chamber FF19 FF20 FF21 PV5 PV6 Locate the drain line at the bottom of each aperture bath Trace the lines through PV (pinch valve) 5 and PV 6 to the flow fittings (FF) on the back panel (FF19 and FF21) The pathway continues in back of the CBC Module and up to the top where the lines come back out through FF3 and FF5 which are located above the waste chamber. Locate FF3 and FF5 Follow the tubing to the waste chamber. Note the check valve present in each tubing. Activity 1. Run a sample and observe the bath draining process SF-15 of 30

126 Preparing the CBC Sample Air pump for diff transfer to sample line Diluent from RBC and WBC diluent dispensers 1. The center section of the BSV rotates to segment three portions of the sample: 1.6 µl for the RBC bath dilution 28.0 µl for the WBC bath dilution 28.0 µl for the Diff dilution 2. The air pump (PM 3)transfers the Diff portion to a sample line just outside (and behind) the BSV. 3. The RBC (PM 11) and WBC (PM 9) diluent dispensers send diluent through the BSV to pick up, dilute and deliver the RBC and WBC segments of blood. Locate the RBC and WBC dispensers in the Main Diluter. BSV (CBC Delivery) Diluent in Diluent in To WBC bath To RBC bath SF-16 of 30 PN CA September 2009)

127 5 CBC lytic reagent pumps Mixing bubble entry 6 4 Dilution entry ports 4. The dilutions are delivered to the aperture baths through the inlet ports on the upper right of each bath. RBC/Plt 10 ml diluent (from RBC diluent dispenser) plus 1.6 µl blood WBC/Hgb 6 ml diluent (from WBC diluent dispenser) plus 28.0 µl blood plus 1.0 ml cbc lytic reagent 5. The CBC lytic reagent pumps (PM 1 and PM 2)send lytic reagent to join the WBC dilution on its way to the WBC bath. This line runs behind the CBC Module. If you begin at the WBC inlet port, follow it back to FF27, find the back side of it and then trace about two inches to a Y-fitting, you will see the smaller diameter tube that comes from the CBC lytic pumps. The WBC/Hgb dilution starts as a cloudy red but becomes a clear red once the lytic reagent reacts with the blood. 6. Mixing bubbles enter both baths from the bottom left port as several bursts of bubbles. The mixing bubbles provide complete mixing of the dilutions ensuring reproducible results. Find the rocket-shaped check valve in the mixing bubble line The check valve is a one-way valve. It lets each aperture bath receive bubbles of gentle pressure (5 psi) Go to the Component Table in the lab to look at an aperture bath, diluent dispenser, a CBC lytic reagent pump and check valves, both small and large. SF-17 of 30

128 Activity 1. Cycle the blood sample. Watch for: BSV Rotating Diluent dispensers making an upstroke. WBC bath for addition of CBC lytic reagent and subsequent clearing of the dilution. (FYI-you can t really see lytic reagent entering the bath since it happens while the diluent and blood are being delivered. All you can see are the effects.) Mixing bubble bursts Self Check 1. Complete the statements about each aperture bath dilution. The RBC/Plt dilution is made up of: µl blood ml diluent The WBC/Hgb dilution is made up of: µl blood ml diluent ml CBC lytic reagent 2. How can you visually tell that lytic reagent has been added to the WBC aperture bath? 3. How could results be affected if no mixing bubbles enter the aperture baths? CBC Sensing System To review The Coulter Principle, refer to the Coulter Hematology Innovations CD, Chapter 1. Each aperture bath has one aperture. Triplicate counting and sizing of cells takes place each time a sample processes, in the form of Count Periods. A count period lasts approximately 4 seconds and is repeated a total of three times. To apply the COULTER Principle for counting and sizing cells, the following are needed to get results from the sample: A suspension of blood cells (specific WBC or RBC dilutions) An aperture of constant size A regulated source of aperture current (passing between two electrodes) A regulated source of aperture vacuum (relative to 6 inches of mercury) SF-18 of 30 PN CA September 2009)

129 The variables in this equation are the actual cells (number and size) of WBC, RBC and Plt of the individual sample. The cell/diluent suspension in the aperture baths: Maintains cell integrity of the cells being analyzed. Conducts electrical current. Aperture Vacuum Low vacuum, equivalent to 6 inches of mercury (6.00 ± 0.04), draws a precise volume of cell suspension from each aperture bath through the aperture. Observations Find the aperture in each aperture bath. Find the Vacuum Isolator Chambers; RBC on the left and WBC on the right. Refer to the CBC module diagram. When the Status is COUNT, low vacuum pulls the dilutions through the apertures to the vacuum isolator chambers. Find the the pinch valves (PV 2 and PV 3) leading from the top of the RBC and WBC baths, respectively, and going to the top of the Vacuum Isolator Chambers. Notice that each I-beam tubing is enclosed by a metal coil. This is used to reduce electronic interference. Find the Low Vacuum Regulator (RG1) Adjustment Knob. Refer to the Main Diluter diagram. The System Test is used if any adjustment to Low Vacuum is necessary. Instruction for using the System Test is in the Troubleshooting Basics (TB) Module. The system counts and sizes cells passing through the apertures. The collected data goes to the Analyzer for further analysis. Activity Go to the Component Table in the lab to look at an aperture bath. Remove an aperture block to see the aperture. 1. Cycle the blood sample. 2. When the Analyzer Status says COUNT, watch droplets coming into the Vacuum Isolator Chambers for RBC and WBC. Self Check How could results be affected by aperture vacuum that is Less than 6 More than 6 SF-19 of 30

130 Aperture Current Aperture current Vacuum Internal electrode External electrode Aperture current Sample beaker Blood cell suspension Vacuum Aperture Aperture tube Aperture housing Bath Internal electrode If an RBC bath, then this is the Sweepflow line. Aperture External electrode Current flows between the internal and external electrodes in each aperture bath. Go to the Component Table in the lab to look at the external and internal electrodes in the aperture bath. SF-20 of 30 PN CA September 2009)

131 Observations Locate the external electrode in each aperture bath. Refer to Figure page 20. The external electrode, a silver-colored, square metal flag, hangs down in the center of each bath. Locate the internal electrodes for each aperture bath. Refer to Figure page 20. A Horizontal cylinder behind and near the top of each bath house the internal electrodes. Aperture vacuum pulls the cells through the apertures. Aperture current is applied between the internal and external electrodes. The cells, poor conductors of electricity, set up an impedance or resistance to the current flow. This impedance, which is proportional to the volume of the cell, allows the cells to be counted and sized. A coaxial cable is connected to the back of each internal electrode. It is through the coaxial cable that information on counting and sizing goes to the Analyzer. Sweep Flow Sweep flow is a flow of diluent behind the RBC aperture during counting. The diluent flow prevents the swirling effect of RBCs behind the aperture It sweeps away the RBCs after they are counted so as not to interfere with platelet counting The sensing zone of the aperture tube extends out from the aperture itself. Without sweep flow, cells (primarily RBCs) tend to swirl or tumble behind the aperture after they are counted. If they nick the sensing zone, they appear as smaller particles and are counted and sized as platelets. Observation Find the sweep flow line below the RBC bath. It has a white plastic sheath around it. Hemoglobin Go to the Component Table in the lab to see the coaxial cables connected to the back of the aperture bath. The hemoglobin measurement occurs after the cell counting completes. Hemoglobin is measured photometrically through the WBC aperture bath. Locate the Hgb Lamp Locate the Hgb pre-amp. SF-21 of 30

132 Preparing the Diff Sample Mixes Erythrolyse II reagent pumps StabiLyse reagent pump The diff blood sample segment is prepared immediately after the two CBC segments process. The BSV rotates back to its original position. The Diff lytic reagent pumps (PM 6 and PM 7) send Erythrolyse II reagent to pick up the 28 µl segment from the line just behind the BSV and deliver it to the mixing chamber. The first rotation of the mixing chamber occurs and the red blood cell membranes are ruptured and dissolved. The StabiLyse pump (PM 12) adds StabiLyse reagent to the mixing chamber during the second mixing process to stabilize the white blood cells and ensure that they remain in a near-native state. Observation Locate the Erythrolyse pumps and Stabilyse pump Locate the mixing chamber Locate the blood/erthyrolyse input port on top of the mixing chamber Locate the StabiLyse input port on the right side of the mixing chamber. Diff Analysis Sample pressure applied to the mixing chamber pushes the prepared Diff sample to the flow cell. At the same time, a sheath stream of diluent from the Sheath Tank is initiated in the triple-transducer flow cell. As the sample stream is pushed into the flow cell, alignment of the sample stream with the sheath fluid (hydrodynamic focusing) allows the white blood cells to pass one at a time through the flow cell aperture. SF-22 of 30 PN CA September 2009)

133 As each white blood cell passes through the Flow cell aperture, the three independent VCS measurements (Volume, Conductivity and Light Scatter) are taken simultaneously. From BSV StabiLyse reagent pump Mixing Chamber To flow cell Activity At the Command Center, change run type to CD. Run a sample and observe the mixing chamber. SF-23 of 30

134 Sample pressure Sheath tank Flow Cell Vented waste Sheath fluid Prepared sample For both Diff and Retic analysis, the instrument injects the sample into the center of a sheath stream for focusing the sample stream into the center of the flow cell aperture. The cells are lined up in single file to go through the aperture for Volume, Conductivity and Scatter analysis. The difference with retics is that the sample and reagents are prepared manually. After this prepared sample aspirates it goes straight to the flow cell for analysis. Locate the following at the Components Table: Laser Lens Block - Focuses the laser on the aperture area of the Flow Cell Flow Cell - Examine the flow cell. Locate the two chambers, the aperture area and the electrodes. Light scatter detector (S reading) Connectors for both the Direct current (V reading) and RF current (C reading) Sample line Go to the Components Table in the lab to see the Triple Transducer Module (TTM) and the flow cells in more detail. SF-24 of 30 PN CA September 2009)

135 Clean Up The Cycle completes Aperture baths cross-rinse with diluent from the diluent dispensers. 10 ml of diluent rinse to the WBC bath comes from the RBC diluent dispenser. 6 ml of diluent rinse to the RBC bath comes from the WBC diluent dispenser. Crossrinse is needed to minimize the potential for carryover in the WBC bath. The WBC bath needs a larger (10 ml) rinse volume to: Remove RBC cell stroma after lysing Remove remaining lytic reagent Rinse above the 7 ml fill line (6 ml diluent + 1 ml CBC lytic reagent) High current is applied to the apertures (the burn circuit) to clean them. The burn circuit activates to remove protein from the apertures. A "crackling" sound indicates that the burn circuit is active. The aspiration system is backwashed and the needle vent is rinsed and dried. The mixing chamber and the sample pathway through the flow cell are rinsed. Waste chambers are drained. Activity 1. Cycle a sample. 2. Watch switching of the rinse in the baths at the end of the cycle. 3. Listen for the burn circuit. Self Check 1. Which bath has more diluent rinse in it at the end of the cycle? 2. Where does diluent rinse for WBC bath come from? 3. Where does diluent rinse for RBC bath come from? 4. The switching of diluent rinse is necessary to clean the WBC bath more thoroughly. List some reasons why the WBC bath needs more clean up. SF-25 of 30

136 Manual Mode Aspiration For the manual mode of operation, you can enter the sample identification by either of the following methods: Scan the bar code label on the tube into the Workstation Command Center Barcode field using the bar code scanner. OR Manually type the bar code number into the Workstation Command Center Barcode field and select <Enter > or <Tab >. Observations In the manual mode, the manual mode aspiration pump draws a maximum of 125 µl through the aspirator tip and into the BSV. An automatic probe wipe washes and dries the outside of the aspirator tip and drains the backwash that cleans the inside of the aspirator tip. Once the blood segment is in the BSV, the cycle continues exactly as in the automatic mode. There is no difference in the handling of the blood dilutions or cycles. Locate the manual mode aspiration pump (PM 5) Locate the Probe wipe Activity 1. Identify a sample to the LH 500 Analyzer. 2. Mix the sample, open the sample tube and immerse the aspirator tip in the sample. Probe Wipe 3. Press and release the aspiration bar. The system aspirates 125 µl of sample. (Remember, the blood detectors are not active in the Manual mode. 4. Be sure to remove and recap the tube when you hear a beep from the instrument. 5. An automatic probe wipe cleans the aspirator tip after sample aspiration. 6. Observe the movement of the probe wipe. 7. Observe the rotations of the mixing chamber for the diff analysis. SF-26 of 30 PN CA September 2009)

137 Answer Key for LH 500 COULTER LH 500 TRAINING MODULES Transport 1. Load stack empty sensor 2. Tube available sensor 3. Tube ram pushes a tube onto the tube forward sensor. Automatic Mode Aspiration 1. Plugged needle vent possible problems: incomplete or partial aspiration. 2. Maximum of 185 µl Sample Delivery 1. RBC dilution: 1.6 µl blood and 10 ml diluent WBC dilution: 28 µl blood, 6 ml diluent and 1 ml CBC lytic reagent. 2. Dilution in WBC bath clears. 3. Results could be erratic on multiple runs of the same sample. Total voteouts could occur. Aperture Vacuum Low vacuum less than 6 : results lower than expected. Low vacuum more than 6 : results on count parameters higher than expected. Clean Up 1. WBC bath has more diluent rinse at the end of the cycle. 2. Diluent rinse for WBC bath comes from RBC diluent dispenser (10 ml). 3. Diluent rinse for RBC bath comes from WBC diluent dispenser (6 ml). 4. WBC bath needs more clean up to: Get rid of cell stroma and protein from the lysed RBCs Rinse above the 7 ml fill line (from the original WBC dilution) Rinse lytic reagent from the WBC bath Completely rinse the WBC bath If you can meet the Objectives of this module to the criteria listed, proceed to the Skill Check. SF-27 of 30

138 SF-28 of 30 PN CA September 2009)

139 SF SAMPLE FLOW SKILL CHECK Analyzer Components Use the Analyzer Component CheckList below. Get numbered tabs from the Skill Check cart. Place the numbered tabs on the correct components. You may work with your partner. When you are finished, call a facilitator to check your work. SCORE: / 24 Facilitator sign off Analyzer Component Checklist 1. Rocker bed 13. CBC lytic reagent pump 2. Tube ram 14. WBC diluent dispenser 3. Tube available sensor 15. RBC Bath mixing bubble check valve 4. Piercing needle 16. Vacuum Isolation Chamber (WBC) 5. Front blood detector 17. Low Vacuum Regulator Adjustment Knob 6. Rear blood detector 18. External electrode RBC bath 7. Auto Mode aspiration pump 19. Erythrolyse pumps 8. WBC aperture bath 20. Sweepflow line 9. Hemoglobin lamp 21. Overflow chamber 10. Waste chamber 22. Mixing chamber 11. Blood Sampling Valve 23. Stabilyse pump 12. Manual Mode aspirator tip 24. Sheath tank SF-29 of 30

140 You may use references as needed to answer the following questions. Sample Dilutions Name the reagents and quantities that make up the following dilutions WBC: µl blood ml (reagent) ml (reagent) RBC: µl blood ml (reagent) Coulter Principle List the four constants necessary for proper application of the Coulter Principle when counting and sizing particles: Name the variable in the application of the Coulter Principle Sample Flow Select any one of the items below and write an explanation of its importance in the diluter cycle. Sweepflow Mixing bubbles Cross-rinse Facilitator sign off SF-30 of 30 PN CA September 2009)

141 SU STARTUP OBJECTIVES Given an operating COULTER LH 500 and access to HELP Locate the STARTUP on the Command Center Initiate Startup cycles on the LH 500 Analyzer Access the Quality Assurance, Daily Checks windows to check results Access the Daily Check Details window Perform the proper procedures in the event of a message on the Daily Checks window, following the HELP procedures Repeat Background from the Command Center Suppress automatic printing of Daily Checks Use (View History) button to view stored Daily Checks results Return to the Daily Checks window using the View Current button Why Is It Important? Quality Assurance, beginning with Daily Checks, is your way of knowing that your LH 500 is in proper working order and ready to run controls and patient samples. Resources To complete this module you will need: LH 500 Analyzer LH 500 Workstation with HELP Laminated Summary Sheets SU-1 of Coulter

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143 INFORMATION / PRACTICE SECTION Startup COULTER LH 500 TRAINING MODULES 1. Make sure power is on at the LH 500 Analyzer and at both the Workstation and Instrument computers. 2. Log on at the Workstation. 3. At a Workstation, go to (HELP) from the Command Center. Go to Search Type Startup Select List Topics Select Performing Daily Startup Click Display 4. Print Performing Daily Startup. 5. For additional information, access the following hypertext topic: Step 4: Check Daily Test Results Print this topic 6. Go back to Search Type Background Select List Topics Select Checking Background Test Results and Display Print this topic (You should now have 3 pages printed. You may wish to staple them together for future reference.) Close HELP. 7. Do you want an automatic printout of Daily Checks? If Yes, go to step 9. If No, go to step Go to Run Configuration now and select the Daily Checks under QA samples. This will automatically print the Daily Checks after a Startup. 9. Select (OK) to save. SU-3 of 6

144 10. Perform the Daily Start-Up following the Help procedure. Select Startup from the Process Type box on the Command Center, then select to begin. Refer to Checking the Daily Test Results to go the screen for results review. Review the results for pass/fail. If any background test fails, perform a background check. Refer to the Help topic Checking Background Test Results to do this background check. If any other subsystem fails (Electronics, Press/Vac, Temperature, Hgb Lamp, Autoloader), select (Daily Check details) to see what failed then repeat Start Up. If you have a pressure or vacuum out of range, then run System Test to adjust the reading. (You will learn more about System Test in the Elective TB-Troubleshooting Basics) Note: When cleaning agent is in the instrument, only a Startup is allowed. 11. Select (View History) to see a list of all previous Daily Checks results. Select a date you wish to view and the results appear on the lower half of the screen. You can also use the button to view the details of these previous results. Practice using this area now to review past startup results. NOTE: If the requested Daily Checks (Startup) results do not appear, more than one line may be selected. After viewing a result, deselect it using the line select button again. The Daily Checks History log stores all Daily Checks results until the next software update or until the workstation is re-imaged. You can delete results from the Daily Checks History log. 12. Select (Current) on the Specific Toolbar to return to the Daily Checks window. Bring printouts from your Daily Checks (Startup) from today, as well as a printout of a past startup (e.g. last week) from the Daily Checks History log to show to a facilitator as part of the Skill Check for this module. SU-4 of 6

145 SU STARTUP SKILL CHECK 1. Show a facilitator the printouts from today s Daily Check (Startup), as well as a printout from the Daily Checks History log of a previous check of at least a week ago. 2. Number the buttons illustrated in the proper order for accessing the current Daily Checks Details window, starting from the Command Center and ending with closing the Details window 3. Show the facilitator how to repeat Background Tests. 4. A message in the Reagent field means you should or. 5. On the LH 500 Workstation, where do you select automatic printing of Daily Checks results? 6. Place a next to the button used to View History of Daily Checks results. Facilitator sign off SU-5 of 6

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147 TB TROUBLESHOOTING BASICS OBJECTIVES Given an operating LH 500, access to HELP, and access to the SF Sample Flow training module: Identify an instrument problem based on Error messages Abnormal sample results Abnormal Daily check or System test results Abnormal control results Isolate a problem based on observation of sample flow Isolate the component(s) involved based on observed symptoms Perform selected cleaning and adjustment procedures as necessary So that an operator can return an instrument to operating condition. Why Is It Important? An operator that can diagnose and correctly respond to simple instrument problems that occur from time to time keeps instrument downtime to a minimum. This saves the laboratory time and money. Resources LH 500 LH 500 Workstation with HELP Diagrams: reference to laminated sheets of instrument modules and components Training Modules: Sample Flow [SF] and Replacing Faulty Components [RFC] Household Bleach be sure to ask facilitator before using Syringe and tubing Cotton-tipped applicators Lint-free tissue Screwdriver TB-1 of Coulter

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149 INFORMATION / PRACTICE SECTION Basic Troubleshooting Techniques How to Recognize an Instrument Problem Start with the data generated: Error messages generated Error messages with corrective actions are listed in HELP. Whenever a message appears on the Workstation, read it and double-click if necessary for more information. Use the History Logs to see if certain messages are occurring very often. Daily Checks Control runs System Test XB Analysis (if used) Sample data IQAP reports Abnormalities in sample flow Steps to More Efficient Troubleshooting Be completely familiar with normal operation: Startup, Sample Analysis, Shut down Normal Sample Flow Know what should happen during each part of the cycle Be familiar with normal sounds of operation Review the SF Sample Flow Module Use a logical approach to obtain a clear symptom and isolate the components involved. Use the error message information as a starting point. Use your knowledge of sample flow Acquire the knowledge and skills necessary to locate and correct problems. Use HELP Use the training modules Observe the instrument Call your Beckman Coulter representative TB-3 of 14

150 TROUBLESHOOTING BY INSTRUMENT SUBSYSTEMS There are three instrument subsystems: Electronic Reagent Fluidic Pneumatics (pressures and vacuums) Hydraulics (liquids) Electronic Subsystem If there is no power: Check to see if the Standby indicator light is on or off Verify position of the Standby/Reset switch, if it is Off, turn it On If the Standby/Reset is On, check the Main Power switch on the back. If it is Off, turn it On. Check all plug connections If all plugs are in and switches are On, check fuses using the HELP procedure If there is a message that Hgb Voltage is low: Check the Hemoglobin Lamp Try adjusting the Hgb Lamp Voltage If it is burned out, replace the lamp using the HELP procedure Reagent Subsystem Level Sensing Two types of sensors provide level sensing: Reagent level sensing uses optical sensors When you see the error message for Reagent Out you must replace the empty reagent container indicated. Always check the indicated container to see if it is empty. If it is empty, replace it. If it is not empty, you have a false reagent out message and may need to call your Beckman Coulter representative. Try priming the reagent first. If you call your BCI rep, you may need to temporarily disable the associated reagent sensor. If a sensor is disabled, make sure a notice is posted warning all operators to visually check the reagent level concerned. To disable a reagent sensor you must: Switch to the Instrument Computer Diagnostics Operator Options Fluidic Test. Select Disable Reagent Sensor press Enter to Begin The Sensor Status Box will display. TB-4 of 14

151 Select with the and arrows on the keyboard the reagent sensor you want to disable. Follow the directions on the screen to complete the task. If a sensor is disabled, the instrument does not know when the container is empty. Sample results will be incorrect. The instrument will not operate until the replacement occurs and you prime the new reagent. Refer to HELP for the Reagent Replacement procedure Waste container level sensing uses a float sensor When you see the error message Waste Full you must replace the waste container. If the waste is not full, check the float sensor, it may be stuck in the up position. The instrument will not operate until the replacement occurs. Refer to HELP for the Waste Container Replacement Procedure. Reagent Related Problems High background counts Reagent may be contaminated. Handle reagents properly to avoid accidentally introducing dust, dirt or powder. Replace contaminated reagent. Dried reagent spills or leaks can form salt deposits that create background electrical noise. Clean all spills and leaks immediately. Expired reagent Be sure to update the reagent replacement screen with the correct dates. Rotate stock to avoid accidental use of expired reagent. Frozen reagent A reagent that was frozen and thawed could have separated chemical components. One symptom could be increased MCVs, due to diluent now being non-isotonic. If there is a possibility of having frozen /thawed reagent (e.g. cold northern climates): If a reagent is still frozen: allow to thaw completely at room temperature. Mix by inversion as per package labeling. Fluidic Subsystem The key components of the fluidic subsystem are the Tubing, Pinch Valves, Pumps and Solenoids. Hydraulics Check tubing connections and routing. Look for leaks or disconnects. At the reagent container TB-5 of 14

152 At the input manifold on the back of the Analyzer Throughout the Main diluter module Check associated pumps, pinch valves and solenoids. Pneumatics There are two main pneumatic signals, vacuum and pressure. Vacuum has two measured levels High Vacuum and Low Vacuum High vacuum is equivalent to inches of mercury. Aspiration, refilling diaphragm pumps and draining aperture baths use high vacuum. It is not operator adjustable An error message alerts the operator of a problem If out of range on the Low side (<17), check the vacuum trap for the presence of liquid (normally it should be dry) and clean as needed. Refer to HELP for the Clean Vacuum Trap Procedure. DO THIS NOW and perform the procedure. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) If the vacuum trap is not the problem, call your Beckman Coulter representative for assisstance. Low vacuum is equivalent to 6 inches of mercury. Derived from the high vacuum Also known as Aperture Vacuum Pulls the dilutions through the RBC and WBC apertures and sweep flow diluent behind the RBC aperture. Operator adjustable An error message alerts operator of a problem Refer to HELP for the Adjust Low Vacuum procedure. Locate the Low Vacuum regulator knob but do not adjust at this time. If the Low Vacuum has been adjusted and it doesn t stay adjusted, check the Vacuum Trap and clean it if necessary. Also check the WBC/RBC VIC s (Vacuum Isolation Chamber) and PV-3 for holes in the tubing or disconnected tubing. Pressure has three measured levels: 60 psi, 30 psi and 6 psi. 60 psi Generated by the compressor in the pneumatic power supply. This is the source of the 30 psi and 6 psi. An error message alerts operator of a problem. If this occurs, call your Beckman Coulter representative. TB-6 of 14

153 30 psi Dispenses reagents from pumps, opens pinch valves and moves air cylinder shafts. Controlled through solenoids. Check solenoid function using the Solenoid Test. Access the solenoid test from the instrument display, Diagnostics Operator Options Solenoid Test. 1. Use the arrow keys to highlight the solenoid you want. 2. Press Enter to select (highlight changes to red text) 3. Press F2-Activate and deactivate. 4. Press F9-Exit DO THIS NOW and perform the procedure. Check the function of Solenoid 7 for the WBC mixing bubbles. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) Operator adjustable An error message alerts operator of a problem Refer to HELP for the Adjust 30 PSI Pressure procedure. Locate the 30 psi regulator knob. Ask a facilitator to verify. DO THIS NOW and then perform the procedure. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) 6 psi Used for Sheath pressure and mixing bubbles Operator adjustable An error message alerts operator of a problem. Call your Beckman Coulter representative before making this adjustment. Locate the Sheath Pressure regulator knob but do not adjust at this time. TB-7 of 14

154 TROUBLESHOOTING THROUGH SAMPLE FLOW To successfully troubleshoot this instrument: Become completely familiar with normal sample flow. Know what should be happening during each part of the cycle Be familiar with normal sounds of operation. This type of knowledge is gained mostly through experience. To increase your comfort level with the normal sample flow of this instrument, periodically review the SF Sample Flow Module. Autoloader Transport Error messages alert the operator of problems with the Autoloader mechanism, sensors, cassette jams. Access the autoloader test functions from the Instrument Computer display (Top, Pos 1). Select Diagnostics Operator Options Autoloader Tests There are six tests that you may use to diagnose problems. Perform the following scenarios to see how these tests work: 1. Select the Right Elevator Up/Down test and observe elevator response. Press Escape. Now do the same with the Left Elevator Up/Down test. 2. Select Rock the Bed. Observe how the mechanism works. Press Escape. 3. Select the Right Elevator Up test and leave the elevator in the up position. Press Escape. Select Autoloader Home and observe what happens. Press Escape when done. 4. Place a cassette in the middle of the rocker bed. Select Clear the Bed/Autoloader Home. Observe what happens. Press Escape when done. 5. Place a cassette in the loading bay. Select Autoloader Test Routine. Observe what happens. Press Escape. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) Cassette problems can be related to buildup up debris or crystals in and on the cassette. Refer to HELP for the Clean Cassettes procedure. If a cassette is jammed and can not be removed, refer to HELP for the Jammed Cassette Removal procedure. DO THIS NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) TB-8 of 14

155 Aspiration Aspiration error messages alert the operator of aspiration problems. The front and rear optical blood detectors determine if the aspirated blood meets the criteria for a proper aspiration. If not, an aspiration error message appears. All sample results flag with a P. In addition, on the Results & Graphics Screen in the Common Area under Status it will state Part Asp. If the blood detectors are faulty, then false error messages occur. Check the blood detectors from the Instrument Computer display (Top, Pos 1). Select Diagnostics Operator Options BSV Tests Blood Detector Test. DO THIS NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) This could be sample related if there is a clot in the tube. Check for a clot before rerunning. It could be a short sample. There must be at least 1 ml of properly collected, anticoagulated blood in the tube for automatic mode aspiration. If sample volume is too low, rerun in manual mode. If the sample has a low hemoglobin (approx. 4 g/dl) the blood detectors post an aspiration error message. This type of sample may be rerun in the manual mode. The needle or aspiration tubing may have a kink, pinch or clog. Check the lines to be sure they are not twisted or pinched. If you suspect a clog in the aspiration pathway, try the Aspiration Path Bleaching (Automatic or Manual) topics in HELP. (Note: Before proceeding, ask facilitator for the fake bleach to use in the classroom). DO THIS NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) As a last resort, replace the needle using the HELP procedure. (this procedure was practiced in RFC Replacing Faulty Components) The BSV may not be rotating properly due to crystalline buildup. Refer to HELP for the Clean Outside of BSV and the Cycle BSV procedures. DO THEM NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) Sample Delivery If the baths do not drain or do not drain completely, the next sample will be over diluted. When overflow is detected in the overflow chamber, an error message appears to alert the operator. Check the drain lines at the bottom of each aperture bath. Be sure they are not pinched. Use a red clip to open the pinch valve and move the bluestriped I-beam tubing back and forth. If the RBC and WBC dispensers are leaking due to cracks or to leaky tubing, too little diluent delivers to the baths. This causes all counts to be too high. Check the dispensers and tubing for leaks. Replace tubing if necessary. Call your Beckman Coulter representative if a dispenser is leaking. TB-9 of 14

156 If the CBC lytic reagent pumps or associated tubing leaks, then too little lytic reagent delivers to the WBC bath. This causes the WBC and the Hgb to be elevated. Observe sample flow for leaks and watch for the color change from cloudy red to clear red in the WBC bath. Replace tubing if necessary. Call your Beckman Coulter representative if a pump is leaking. If the mixing bubbles are missing from either bath, this can cause erratic results and increased CP voteouts. Check the bubbles by observing sample flow or from the Instrument Computer display (Top, Pos 1),. select Diagnostics Operator Options Fluidic Tests Bubble Mix. DO THIS NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) Verify the mixing bubble check valves are not leaking or plugged. Replace if necessary. CBC Sensing If the aperture vacuum or Low Vacuum is out of range, an error message alerts the operator of the problem. Use the System Test to adjust Low Vacuum. If either aperture becomes clogged or partially clogged, there could be CP Voteouts (V, v) or Total Voteouts (-----) of the measured parameters. If this happens on just one sample, repeat it. If this begins happening on most or all samples, from the Instrument Computer display (Top, Pos 1), select Diagnostics Operator Options Fluidic Tests Multiple Aperture Zap. Refer to the HELP procedure Zap Apertures. DO THIS NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) Multiple Aperture Zap may be done more than once. It usually cures the problem very quickly and easily. If the voteout problem persists after trying the previous procedure, then you can try bleaching, but this should only be done as a last resort because it is time consuming. Refer to the HELP procedure Bleach Apertures and Flow Cell. DO THIS NOW. (Note: Before proceeding, ask facilitator for the fake bleach to use in the classroom). Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) If the Hemoglobin Lamp voltage is too low, an error message alerts the operator. Refer to the last section of the HELP procedure Replace Hemoglobin Lamp Assembly to find the Hemoglobin Lamp Adjust procedure. If this fails, replace the lamp. (Both of these procedures were practiced in the RFC Replacing Faulty Components module) TB-10 of 14

157 Diff or Retic Sensing If there is a flow cell problem, one of three messages appears: PC1, PC2 or FC along with the code (:::::) in place of results. PC1 could be caused by: A partial obstruction of the sample line from the mixing chamber to the flow cell. A partial flow cell obstruction. Flow cell sample or sheath pressure errors (This generates the specific pressure error message at the instrument and Workstation) PC2 could be caused by: Insufficient Diff reagent delivery Patient chemistry imbalance (common with sample problems such as elevated triglycerides, improper sample collection/mixing, or drug interferences, i.e. chemo) Mixing chamber not rotating FC could be caused by: Full clog of the flow cell. The instrument tries Autoclearing the flow cell, but if unsuccessful, the system stops. Not all flow cell problems are clogs. They may be related to an individual sample. From the Instrument Computer display (Top, Pos 1), select Diluter Functions Purge. DO THIS NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) If Purge does not work, try running Latron. Run the Latron Primer as usual, if OK proceed to next step. Run the Latron Control as if it were the Primer In other words, keep all of the primer settings on the Command Center, but run the Control. If the Flow Cell is clear, you should recover the number DO THIS NOW. Have your partner signoff on the Skill Check. (If you do not have a partner, check mark the Skill Check.) This means there is no clog and it must be another problem (see next step). If you recover a very low number, then either the sample line from the mixing chamber to the flow cell has a clog or the flow cell has a clog. Try the Bleach Apertures and Flow Cell procedure using HELP. If the flow cell is not clogged, then it could be a VCS Technology problem, a chemistry problem or a mixing problem. VCS Technology problem: Run Latron Control the correct way. If all values recover within limits, go to next step. If not, call your Beckman Coulter representative. Chemistry problem: if the sample is very lipemic, do the diff manually. If the problem appears on many samples, consider the LH Series Pak reagents. Be sure they are not expired. Check the respective reagent pumps and tubing for leaks. Replace if necessary. TB-11 of 14

158 Mixing problem: observe the mixing chamber rotation during sample flow. If the chamber is not rotating, call your Beckman Coulter representative. Proceed to the Skill Check for this module TB-12 of 14

159 TB TROUBLESHOOTING BASICS SKILL CHECK Instrument Problem Recognition 1. Name at least four types of data that may be used to recognize an instrument problem. 2. What are the three main steps to more efficient troubleshooting? 3. What are the three instrument subsystems? Aspiration Error Messages 1. List at least three possible reasons for Aspiration Errors: 2. What Aspiration Error message appears under STATUS when you run blood samples with the blood detectors disabled? 3. How does the Workstation flag sample results when there is any Partial Aspiration situation? Diff and Retic Error Messages and Flags 1. List one reason for the occurrence of each of the following messages or flags: PC1 PC2 ::::: TB-13 of 14

160 Troubleshooting Diff and Retic 1. Given the following scenarios what problem(s) might you suspect, if any? LATRON Control run as a Primer = 8192 LATRON Control run as a Primer = 0 Colleague Signoff of Procedures Performed or Check-Off if Solo Clean Vacuum Trap Blood Detector Test Use the Solenoid Test to observe function of Sol. 7 Aspiration Path Bleaching (Automatic and Manual) Adjust the 30 psi Clean Outside of BSV Right Elevator Up/Down Cycle BSV Left Elevator Up/Down Bubble Mix Rock the Bed Multiple aperture Zap Autoloader Home Bleach Apertures and Flow Cell Autoloader Test Routine Purge Jammed Cassette Removal Troubleshooting by Symptoms Run Latron Control as if it were Latron Primer 1. You observe that the past three results all have a very high WBC and Hgb. What would you suspect? Where would you look? What could you do? 2. You observe that you have an increased number of CP voteouts on RBC and Plt results. What would you suspect? Where would you look? What could you do? 3. Your low vacuum won t adjust. What would you suspect? Where would you look? What could you do? Facilitator sign off TB-14 of 14

161 WB WORKSTATION BASICS OBJECTIVES Given an operating COULTER LH 500 System, be able to: Identify the Instrument Computer and the Workstation by pointing to each Tell whether a display is from the Instrument Computer or the Workstation describing each to a facilitator Switch between the two computer displays using the Computer switch box Log on and log off the Workstation Identify the icons of the Command Center by labeling a picture Use Tool Tips and know why they are used Shutdown and restart the Workstation to address error situations Navigate through the Instrument Computer screen using the keyboard following written instructions Reboot the Instrument Computer using the Standby/Reset switch Define the traffic light colors Use the on-line Help to find information using the Contents, Index and Search features and print topic information Be able to use Help Mode button Create Favorites Know where to set up the Automatic To Do List Deletion feature Why Is It Important? The LH 500 Workstation uses the Microsoft Windows 2000 Operating System. The skills contained in this module are common to most Windows-based programs and will help you in other Windows encounters! The LH 500 Instrument Computer uses an MS DOS-based system. The skills in this module will familiarize you with navigating via the keyboard. The LH 500 HELP gives you all your instrument manuals on-line with easy access to many features unavailable in print. With practice you will see that using HELP is a convenient way to access information that is right at your fingertips Resources LH 500 Hematology Analyzer WB-1 of Coulter

162 INFORMATION / PRACTICE SECTION LH 500 System The LH 500 System includes: The Analytical Station Analyzer module, Diluter modules and Power Supply. The Analytical Station is also referred to as the Instrument. The LH 500 Workstation Computer a Windows 2000 operating system and display with keyboard and mouse interfaces. The LH 500 Instrument Computer an MS-DOS based system that shares the same monitor display and keyboard with the Workstation. A computer switch box - allows selection between the two computer bases and determines the display you see on the monitor. A digital communication box allows communication between the Instrument Computer and the Workstation Computer. This learning module covers the two computers and the switch box. Computer Switch Box Light indicates which position is currently in use. Press button to change displays. Top (Pos 1) Bottom (Pos 2) The switch box has two buttons used to select which computer display you need to see. The two computers share the same monitor and keyboard. The top switch (Position 1) connects to the Instrument Computer, the monitor and keyboard. The bottom switch (Position 2) connects to the Workstation Computer, the monitor, the keyboard and the mouse. Note:You may have a different type switch box. See facilitator for demo. WB-2 of 22

163 There are two ways to switch from one display to the other: Simply press the top button or the bottom button. Use the keyboard Press the Scroll Lock key twice (you will hear a beep) and then the Up arrow key or the Down arrow key. How to Tell Which Display You Are On: The Instrument Computer display has a blue background and has three main menu headings across the top: Diluter Functions---Setup---Diagnostics. Use the keyboard arrow keys to highlight your selection and then press the Enter key. The Workstation Computer display has many different windows-type screens. If the logon screen appears then a user name and password must be typed. This should be the case right now if you are on this display. If the monitor display is blank, try pressing the spacebar twice to bring it back. If that doesn t work, try moving the mouse a bit. Try This Look at your monitor screen now. Bring up the display, if blank. Now look at the display to see which computer screen appears. Use the switch box to change to the other display. Be sure you can tell the difference. If not, please ask a facilitator. Press the bottom switch when you are done to be at the LH 500 Workstation. LH 500 Workstation Log On to Windows 2000/LH 500 program When the Workstation is logged off or if it was Shutdown and then Powered off/on, the Log On to Windows screen appears. Both a User Name and Password are required: For ease of use on all LH 500 Workstations in the classroom, the User Name is Labadmin (not case-sensitive). WB-3 of 22

164 For class purposes the Password is coulter (lower case). Passwords are case-sensitive. Type in the password now and press Enter from the keyboard. NOTE: The password coulter is not installed on the Workstation in your laboratory. Upon installation, the password for LabAdmin is blank (no password required). You may select your own password. The LH 500 Workstation computer system supplied with your instrument includes: an Intel 1 GHz Pentium III processor with 256 MB RAM, 40 GB Hard Drive, CD-RW, DVD/CD-ROM, 3.5-inch diskette drive, optical mouse and 17-inch Flat Panel LCD display monitor with built-in speakers. Let s look at the Workstation desktop display next and learn some of its terminology. LH 500 Workstation Display Windows 2000 Desktop Windows 2000 Desktop Icons Command Center Instrument Status WB-4 of 22

165 Desktop Icons COULTER LH 500 TRAINING MODULES The five desktop icons on the upper left are functional windows icons. However, it is best not to use them. They are not necessary to operate the LH 500. All functions needed by the operator are accessible through the Command Center. Command Center The Command Center is the green bar located across the bottom of the desktop area. Think of the Command Center as the Main Menu of the Workstation. The user currently logged on. Process Type Usually has Auto Analysis selected Run Type Select the test mode to run in Asp Mode Auto (closed-vial) or Manual (open-vial) Traffic-light icon Instrument Message Status Eight LH 500 screen application buttons Stop/Start Used to stop or start selected processing configuration Barcode entry Scanned or Typed Enable/Disable Blood Detectors Select # aspirations/tube Predilute Option with dilution factor entry A ToolTip appears over a Workstation button when your cursor moves or flies over the button. It remains on screen for a few seconds to tell you the name of the button. Try this now. Hold the cursor over a button without clicking. Quality Assurance When you move the cursor towards the bottom of the screen, the Windows Taskbar may appear. Just move away from the taskbar to make it disappear, and then move back to the Command Center make your selection. Try this now. Windows Taskbar Instrument Status The Instrument Status box shows the status of the Instrument Computer. Some examples: SYS NOT READY Instrument is in a Clean or Shutdown cycle SELECT FUNCTION Instrument is ready but Command Center choices are not started AUTO MODE ON Instrument is in the Automatic (closed-vial) mode of operation MANUAL MODE ON Instrument is in the Manual (open-vial) mode of operation WB-5 of 22

166 COMPRESSOR OFF the compressor timed out (after one hour of non-use) or was turned off STARTUP instrument running the startup cycle and tests The same message displays on both the Instrument Computer (at the lower right) and the Workstation Computer. LH 500 Screen Application Buttons Use your mouse to view the tool tip name of each of the LH 500 screen application buttons below. Read the descriptions on the next page to familiarize yourself with what information is found in each area. Select each button on the Command Center now to view the area that opens. WB-6 of 22

167 When you want to change the area you are viewing, just select a new button from the Command Center The program automatically closes what was open and then opens the new choice. There are three exceptions: If the Run Configuration screen is open, you exit by selecting either (OK, save and close) or the red to cancel any changes you made. If the Help screen is open, you exit by selecting the small x in the upper right corner. If the History Logs are open, exit by selecting the small x in the upper right corner. Asp Mode Field Use the drop-down box to choose the aspiration mode for running samples. Auto is the closed-vial mode and Manual is the open-vial mode. The field defaults to Manual if the Run Type is R or if Process Type is Control and Run Type is Ltx Prime or Latex. Stop and Start Buttons Select the Stop button whenever you want to change the Process type or Run Type or any of the choices on the Command Center. In fact, anytime that any choice needs to be changed, you must select Stop first. Select the Start button after making your selections on the Command Center. You must press start after making your selections or the instrument will not respond. Predilute and Factor Fields The LH 500 allows you to run dilutions of samples with very high counts. After you make your sample dilution, select the check box for Predilute. The Enter Dilution Factor window appears as shown below. Try this now. You may enter your dilution multiplication factor from 1.1 to 5.0. Sample dilutions may only be run in the Manual processing (aspiration) mode and use only the CBC test mode. The factor entered remains in effect for one sample only. It then reverts to 1.0. WB-7 of 22

168 Traffic-light Icon The traffic light icon shows red, yellow or green lights to indicate the instrument s status. A green light means all is well; there are no errors or messages. A yellow light means the instrument recorded a message, but it is information only. The instrument is still operating. You should look at the message when you have a chance (double-click on the yellow light) and then acknowledge it. A red light means the instrument stopped due to an error or malfunction or Auto-Stop request. An audible alarm sounds. Double-click on the red light to read the message(s) and then acknowledge it. WB-8 of 22

169 Log Off The Workstation COULTER LH 500 TRAINING MODULES Select from the Command Center. The Log Off The System window appears. The default selection is Log Off Current User. The other choice is Shutdown the Workstation. Choose this to close the operating system properly and power off the computer. Choose the default to change the current user logged on to the system (e.g. at the end of a shift). OK to Log Off or Shutdown Continues with selected option. Cancel Closes window and returns to Desktop. Context-sensitive Help on the Shutdown topic. Shutting Down the Workstation NOTE: The Workstation should be powered down at least once a week for the Database to update and reallocate memory. A good time to do this would be during the Clean or Shutdown cycles of the Analyzer. In addition to the above recommendation, you may have to shut down and restart the Workstation in response to certain error situations or if the computer locksup or doesn t respond. Be sure to read any Help message for error recovery carefully for this indication. In the event of a communication problem between the Workstation and the LH 500 Analyzer, you may need to reset the Analyzer using the RESET button on the lower left side of the Analyzer cover as well as selecting to shut down and restart the Workstation. Look up in HELP under the SEARCH TAB the topic System Lockup and read it. Practice 1. Be sure you are on the Workstation display and select from the Command Center. WB-9 of 22

170 2. Select Shutdown the Workstation. Select OK. 3. The Workstation will save any unsaved data and display a window notifying you it is shutting down the workstation. Next a window is displayed that states you may now power off the workstation. 4. Turn Off the power using the power button on the front of the LH 500 Workstation computer tower. 5. After one minute, turn the power back on. Log on to the workstation when the Log On window appears. LH 500 Instrument Computer Main Menu You switch from the Workstation Computer to the Instrument Computer only to access certain Diluter Functions and Diagnostics. Press the top button at the switch box to change to the Instrument Computer display. There are three Main Menu choices: Diluter Functions Setup Diagnostics Diluter Functions Setup Diagnostics Prime Reagents Disinfect Clean Rinse Purge Diluent Lyse Pak Cleaner All Set Date/Time Optimize Hard Disk Requires Service Diskette and Password Multiple Aperture Zap Compressor On/Off Disable Reagent Sensors Bubble Mix Clean Needle Cycle BSV Blood Detector Test Probe Wash BSV Removal Autoloader Home Clear the Bed/Autoloader Home Rock the Bed Right Elevator Up/Down Left Elevator Up/Down Autoloader Test Routine Operator Options Service Options Fluidic Tests BSV Tests Autoloader Tests System Test Solenoid Test Drain and Vent HGB Lamp Adjust Enter Calibration Factors Copy Raw Data to Diskette WB-10 of 22

171 Navigate through the menu tree using the Keyboard only (the mouse will not work in this interface). Use the Arrow keys to move Left, Right, Up or Down. The menu selections are highlighted. Once highlighted, press the Enter key to choose it. Many times you must press Enter a second time to begin the function. Just read the screen and follow the directions given. To back out of a menu choice simply press Esc or the F-9 key. Notice that each of the menu choices on screen have one letter highlighted. This is an alternate way to navigate through the menu. Simply press the keyboard letter corresponding to the highlighted letter. This eliminates the need to press Enter to choose the selection. However, you will still need to press Enter when requested on the screen. Diluter Functions Prime Reagents use this function whenever you replace a box or cube of reagent. Before you prime, remember to enter the required lot information in the Workstation Reagent setup screen. Disinfect use this function to prepare the instrument for shipping and to bleach the apertures and flow cell. Clean this is the desired function for the daily Clenz shutdown for the instrument. It automatically performs Startup 30 minutes later. Rinse use this function in conjunction with troubleshooting bath problems. It does a Drain and Rinse (refill) of the two baths and the flow cell. Purge use this function to clear a possible flow cell clog. It purges the flow cell with Clenz and then returns to sheath (diluent). Setup Set Date/Time use this function to set the date and time the instrument uses to timestamp sample results. If you are in an area that follows Daylight Savings Time, you need to change the time twice a year. Optimize Hard Disk this utility begins automatically any time the instrument reboots (Standby/Reset). Use this function to begin the utility at any other time. Diagnostics Operator Options use these functions for troubleshooting. You will be introduced to each of these functions in the Troubleshooting Basics module. Service Options these functions are only available to your Beckman Coulter representative. Status Along Bottom of Display F9-Exit CtrlF9-Stop CtrlC-Clear the Bed If the Status is RED with an error message, use Alt-End to stop the beeping. WB-11 of 22

172 Practice Try moving around the menu: Highlight Diluter Functions and press Enter. Highlight Prime Reagents and press Enter. Press Esc twice to get back to the Main Menu. Now try the alternate way: Press i (to choose Diagnostics) Press O (to choose Operator Options) Press B (to choose BSV Tests) Press B (to choose Blood Detector Test) (Do not press Enter at this time!) Press F-9 (to choose Exit) Standby/Reset Switch Locate this rocker-type switch on the lower left side of the instrument. Use this switch to place the instrument in a Ready or a Standby state. In the Ready state, the back portion of the rocker switch is flush with the instrument s base; this is position I. This corresponds to the green LED on the front of the instrument. In the Standby state, the front portion of the rocker switch is flush with the instrument s base; this is position O. This corresponds to the green LED on the front of the instrument. In the Standby state, voltages are still being applied to a memory location in the Analyzer, but everything else is powered down. Use the switch to reset the system or reestablish communication between components. To reset the system, place the Standby/Reset switch in Standby (position O) for 15 seconds, and then flip the switch back to Ready (position I). The Instrument Computer goes through a reboot process. The optimize program starts automatically to perform daily, weekly or monthly scans of the hard drive. This will only happen once on any day, so if a reboot happens twice, it skips the optimize process the second time. WB-12 of 22

173 If you want to cancel the optimize program press Esc and then answer yes Y to the question Do you want to cancel optimize? Y/N The Instrument Computer should optimize once a week. If you have not reset the system during the week, you should use the standby/reset switch to reboot and optimize. If you observe that BOTH of the LEDs are off, this means that either the Main Power switch on the back of the instrument is OFF or that the instrument is unplugged or a fuse is blown. The red button with the two vertical white bars is the Emergency Stop. Only use this to immediately stop the instrument if something is going very wrong (e.g. a tube breaking). Do Not Touch This Button in Class. Practice Place the Standby/Reset switch in Standby (position O) for 15 seconds, and then flip the switch back to Ready (position I). Observe that the Instrument Computer reboots. Note: You do not have to be at the Instrument Computer display to use the Standby/Reset Switch. We just want you to be familiar with what happens when you are! When the reboot finishes, you should be back at the Main Menu and the Status in the lower right of the screen should display SELECT FUNCTION. It may also say SYSTEM NOT READY if the unit is in a Shutdown cycle. Press the bottom switch at the switch box or use keyboard commands to return to the Workstation display. Note that the Instrument Status box also displays SELECT FUNCTION. Review the System Lockup topic in HELP. WB-13 of 22

174 The LH 500 HELP System The LH 500 program has an extensive, multimedia HELP system built-in. When all other windows (screens) are closed, selecting the Help button from the Command Center opens the following screen. Do this now. The Help screen uses a format similar to internet browsers such as Internet Explorer and has three main areas: Browser Menu Bar Buttons Help Window Navigation Pane Help Topic Display Area WB-14 of 22

175 Browser Menu Bar Buttons Use the buttons located here to navigate Back and Forward (through screens you have just viewed), Print, Go to a Glossary, etc. Three small buttons located in the upper right corner are used to Minimize, Maximize (or Restore Down) or Close the Help screen. Minimize Close Restore Down Maximize Help Topic Display Area The right side of the screen displays the topic chosen. When you choose the Help button from the Command Center, it always opens to the topic Using Command Center. Help Window Navigation Pane This area has four tabbed sections and may be hidden by selecting the Hide button on the browser menu bar. Use the Show button to redisplay the navigation pane. (Try this now!) Each section provides a different way of accessing or finding the Help topic you want to read. Whatever topic is viewed in the Topic Display Area will also be highlighted in blue in the Navigation Pane Area. Contents used as a Table of Contents, shows broad subject areas. Index type a keyword in the white box near the top and the index scrolls to the possible topic choices. If you see the topic you want, there are two ways to open it: double-click on it or highlight the topic and then click on the Display button at the bottom of the screen. If you have Help maximized, this Display button is hidden behind the Command Center. In this case, click on the Restore Down button in the upper right corner (the middle button). WB-15 of 22

176 Search similar to Index. Type a word or words in the white box near the top, then select the List Topics button or press Enter. If the topic you are looking for appears in the list, select it and then select the Display button. When your topic appears, notice that the word or words that you did the search on are highlighted wherever they appear in the topic. The three checkboxes at the bottom of this screen also influence search results. Try having different boxes checked and search again. If the topic name is cut off, you can click on the vertical bar and drag to the right or double-click and it will resize to the longest title. Vertical Bars Using Contents Favorites this tab allows you to bookmark topics that you refer to over and over, so you do not need to look them up each time you open Help. Once you have the particular topic open, select the Favorites tab and then select the Add button at the bottom of the screen. The next time you need your topic, open Help, select the Favorites tab and choose the topic you want to view. 1. Log on to your individual Workstation. 2. Select from the Command Center The Using Command Center HELP window appears. WB-16 of 22

177 3. Select the hypertext jump (blue text) Buttons that will take you to a topic description of Workstation Command Center Buttons. You may print any HELP topic from the Workstation. 4. Single-click with the left mouse button on the Contents tab. Using Help Single click on the or double click on the words Using Help. Using Help (topic) Single click on the or double click on the words Using Help. 5. Click on Print (Help Menu Bar) to print this topic. The Print Topics window opens. Choose Print the selected topic. Next the Print Dialogue window opens. See next page for information. NOTE: This screen is for example only. DO NOT change the printer already chosen in class. Also, depending on the printer, this screen may vary slightly in appearance. The printer you are using should be highlighted automatically on the General Tab. If you want more than one copy, type in the number you want on the General Tab. Choose the Setup tab to change the orientation of printout to Landscape. This option may be in a different tab. It depends on the printer you have. Select Print 6. At the top of the HELP window, find the Back button. Select Back. 7. Where did the Back button take you? 8. From Using Command Center select Process type field (hypertext jump) Now find another topic using CONTENTS WB-17 of 22

178 9. In Contents, click on Using Index Quick Reference System Components LH 500 Front View 1. Click on the Index tab 2. Type LATRON. The topic LATRON is highlighted. 3. Click on. (Reminder: if Display is hidden, choose the Restore Down button or simply double-click the topic LATRON.) 4. Select Running Latex (Control) Diff and Retic. 5. Click on 6. Print the topic and save it for use in the Quality Control [QC] module. Using Search 1. Click on the Search tab. 2. Type calibration. 3. Click on List Topics. How many topics are found? 4. Place the cursor on the vertical line between Title and Location and then click and hold the left mouse button and drag the arrow to the right until you see complete titles for all topics. 5. Select Calibration Overview, then or double-click. The opened topic has the word calibration and other related words highlighted in blue. This is unique to Search. 6. Print the topic and save it for use in Calibration [CAL] module. Help Mode 1. Close HELP by clicking on the (Close button) at the upper right corner of the window. 2. On the Workstation Command Center, select (Patient Results), the Patient Tests [Results & Graphics] window displays. If the Results & Graphics view does not display, select the button from the common toolbar. 3. Select the Parameters tab. 4. With the left mouse button, click on the (Help Mode) button on the Common Tool Bar (at the top of the screen). 5. Release the mouse button. 6. Move the cursor (with question mark) to the WBC field. WB-18 of 22

179 7. Click on the WBC field. A pop-up window opens. Help Mode gives you additional information about on-screen items without having to access a HELP topic. 8. Close the Help pop-up window by selecting the. NOTE: It is always necessary to close the Help window when finished or it may become stuck on another window. If this occurs, go to the Run Configuration screen and you will see the Help screen behind the Run Configuration screen. Click on the Help screen and close it. Creating Favorites You may add any topic from Help to the Favorites list. Begin by finding the topic you want to add to the list and open it. Once opened, click on the Favorites tab and you will see the topic title near the bottom. Click the Add button. Use the Contents, Index or Search tabs to locate the following topics, and then put them in the Favorites tabbed area: Replace Reagent Containers Running Latex Control Diff and Retic Cycling CBC/Diff Control in Automatic Aspiration Mode Message List Topics may be Added or Removed as necessary. Try this now. WB-19 of 22

180 Automatic ToDo List Deletion Step 1 Action At the Workstation, from the Command Center, go to Setup) (Database Preferences) (System 2 Under Automatic ToDo List Deletion, select the Delete ToDo List Entries. 3 Select the time for deletion. [Default setting is Older than: 24 hours.] It can be set anywhere from Hours. Do not save at this time as we do not want to delete the classroom ToDo list. Select. 4 Be sure you understand all the terminology and practice the skills mentioned in this entire module. Take as long as you need to have confidence in this information. When you are confident in your ability, proceed to the Skill Check for this module. WB-20 of 22