COULTER LH 500 HEMATOLOGY ANALYZER

Transcription

1 COULTER LH 500 HEMATOLOGY ANALYZER TRAINING MODULES This document is not intended to replace the information in your instrument Instructions for Use manual (IFU). Information in the Instructions for Use manual supersedes information in any other manual.

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 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 into your computer. Only operate your system s computer with software authorized by Beckman. 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 or an authorized Beckman distributor, and, if it is not presently under a Beckman service maintenance agreement, Beckman 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 Representative. ii of iv

3 TRADEMARKS 5C Series Control, Beckman,, Counter, Lin-C, Lyse S, S-Cal, Z Series are trademarks of Beckman, Inc.; Beckman, the BECKMAN COULTER logo, COULTER, Counter, 5C Series Control, Lin -C, LYSE S, S-CAL and Z Series are registered in the USPO. All other trademarks, service marks, products or services are trademarks or registered trademarks of their respective holders. Copyright Beckman, Inc All Rights Reserved iii of iv

4 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 Revision CA.1 (June 2012) Software Release 2A4 iv of iv

5 COURSE GUIDE Implementation Troubleshooting Setup Basics This document is not intended to replace the information in your instrument Instructions for Use manual (IFU). Information in the Instructions for Use manual supersedes information in any other manual. 1 of 6

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7 TRAINING TOPICS BASICS SETUP TROUBLESHOOTING IMPLEMENTATION 3 of 6

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9 GROUP DISCUSSIONS The following discussions take place in the classroom/laboratory. Ending times noted are approximate. Tuesday General Information Learn about each other Safety: Personal Protective Equipment and Fire Evacuation Introduction to the course and materials Location of additional reference materials First day Registration Questionnaire Wednesday Review of Patient Result Screens Review of QA area Thursday Friday Flags, Codes and Messages Review PROService and Intelligent On-line Service Info Overview of Key Operator Certification program Overview Beckman website Last day self-evaluation, course evaluation and instructor evaluation Graduation 5 of 6

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11 BASIC OPERATION MODULES Implementation Troubleshooting Setup Basics This document is not intended to replace the information in your Instrument Instructions for Use Manual (IFU). Information in the Instructions for Use Manual supersedes information in any other manual. B-1of 68

12 You will see the following symbols used throughout the learning modules. Important information you should know. Techie-type information you may be interested in knowing. Do these steps now! Timely Tip You reached the goal! I do not understand Questions Resources, Help B-2 of 68

13 COURSE MAP - BASICS Begin Here Inovations Prerequisite Workstation Basics Startup Quality Control Running Samples Shutdown B-3 of 68

14 Name PERSONAL PROGRESS SUMMARY Basic Operation Modules Date Initial Facilitator Innovations Prerequisite Workstation Basics Startup Quality Control Running Samples Shutdown B-4 of 68

15 WB WORKSTATION BASICS OBJECTIVES Given an operating COULTER LH 500 System, be able to: Identify the Instrument Computer and the Workstation Tell whether a display is from the Instrument Computer or the Workstation 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 Use Tool Tips and know why they are used Shutdown and restart the Workstation Navigate through the Instrument Computer 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 Be able to use Help Mode button Create Favorites 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 B-5 of 68

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17 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 bottom switch (Position 2) connects to the Instrument Computer, the monitor and keyboard. The top switch (Position 1) 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. B-7 of 68

18 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. 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). For class purposes the Password is coulter (lower case). Passwords are case-sensitive. Type in the password now and press Enter from the keyboard. B-8 of 68

19 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. 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 Desktop Icons 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. B-9 of 68

20 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 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. B-10 of 68

21 Select each button on the Command Center now to view the area that opens. 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. B-11 of 68

22 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. 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. B-12 of 68

23 Log Off The Workstation 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 the default to change the current user logged on to the Choose this to close the system (e.g. at the end operating system of a shift). properly and power off OK to Log Off or Shutdown Continues with selected option. Cancel Closes window and returns to Desktop. Context- sensitive Help on the Shutdown topic. B-13 of 68

24 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 Step Action 1 Be sure you are on the Workstation display and select from the Command Center. 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 B-14 of 68

25 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 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). B-15 of 68

26 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 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. Practice Try moving around the menu: Step Action 1 Highlight Diluter Functions and press Enter. 2 Highlight Prime Reagents and press Enter. 3 Press Esc twice to get back to the Main Menu. Now try the alternate way: Step Action 1 Press i (to choose Diagnostics) 2 Press O (to choose Operator Options) 3 Press B (to choose BSV Tests) 4 Press B (to choose Blood Detector Test) (Do not press Enter at this time!) 5 Press F-9 (to choose Exit) B-16 of 68

27 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. 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. B-17 of 68

28 Practice Step Action 1 Place the Standby/Reset switch in Standby (position O) for 15 seconds. 2 Flip the switch back to Ready (position I). NOTE Observe that the Instrument Computer reboots. 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. 3 Press the bottom switch at the switch box or use keyboard commands to return to the Workstation display. NOTE The Instrument Status box also displays SELECT FUNCTION. Review the System Lockup topic in HELP. B-18 of 68

29 The LH 500 HELP System COULTER LH 500 TRAINING MODULES 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 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 B-19 of 68

30 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). 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. B-20 of 68

31 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 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. B-21 of 68

32 Using Contents Step Action 1 Log on to your individual Workstation. 2 Select from the Command Center. The Using Command Center HELP window appears. 3 4 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. 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. Click on Print (Help Menu Bar) to print this topic. The Print Topics window opens. Choose Print the selected topic. 5 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. B-22 of 68

33 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 Select Back again. 9 From Using Command Center select Process type field (hypertext jump) 10 Now let s find another topic using CONTENTS In Contents, click on Quick Reference System Components LH 500 Front View Using Index Step Action 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. B-23 of 68

34 Using Search Step Action 1 Click on the Search tab. 2 Type calibration. 3 Click on List Topics. How many topics are found? 4 5 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. 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 Step Action 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. B-24 of 68

35 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 Step 1 2 Action 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. 3 Click the Add button. 4 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. 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. B-25 of 68

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37 WB COULTER LH 500 TRAINING MODULES WORKSTATION BASICS SKILL CHECK 1. Identify these parts of the Command Center: 2. Define the traffic light colors, what they mean, what you should do: Green light Yellow light Red light At any Individual Workstation with a facilitator present: 3. Be sure you are on the Workstation display. 4. Should the Workstation be Shutdown routinely? If yes, how often and for what purpose? 5. Show the facilitator how to Shutdown a Workstation. 6. Name one time you may need to Shutdown a Workstation. 7. Go to an Instrument-connected Workstation and change to the Instrument computer display. Name the three Main Menu choices: B-27 of 68

38 8. Demonstrate the two ways of moving around the Instrument Display menu to your facilitator. 9. Be prepared to demonstrate the following to your facilitator: o Show the menu path to prime the CBC Lyse. o Show the menu path rock the Autoloader bed. 10. Reboot the Instrument Computer using the Standby/Reset switch. Relate the meaning of the green LEDs on the front of the instrument to this switch. Using HELP MODE 1. At the Workstation, go to (Patient Results). You should be on the Patient Tests [Results & Graphics] window. 2. Click on the CBC Data tab. 3. Use the HELP MODE button to find additional information about the RBC Histogram. 4. From the RBC Histogram HELP window displayed, answer the following question: 5. How do you view an enlarged version of a histogram? 6. Close the HELP window. Creating Favorites 1. Open HELP and show your facilitator the Favorites you added. 2. Demonstrate how to delete Favorites. B-28 of 68

39 SU COULTER LH 500 TRAINING MODULES 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 B-29 of 68

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41 INFORMATION / PRACTICE SECTION Startup COULTER LH 500 TRAINING MODULES Step Action 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. At a Workstation, go to (HELP) from the Command Center. 3 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 8. 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. B-31 of 68

42 9 Select (OK) to save Performing a Daily Startup Step Action 1 Perform the Daily Start-Up following the Help procedure. 2 Select Startup from the Process Type box on the Command Center, then select to begin. 3 Refer to Checking the Daily Test Results to go the screen for results review. 4 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 5 Select (View History) to see a list of all previous Daily Checks results. 6 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. 7 Select a Startup from at least a week ago and print it. 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. 8 Select (Current) on the Specific Toolbar to return to the Daily Checks window. B-32 of 68

43 Show 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 the facilitator as part of the Skill Check for this module. B-33 of 68

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45 SU COULTER LH 500 TRAINING MODULES 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. 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. B-35 of 68

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47 QC COULTER LH 500 TRAINING MODULES QUALITY CONTROL OBJECTIVES Given an operating COULTER LH 500 Series Analyzer, Instructions for Use for 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 Instructions for Use and HELP procedure Access control folders using the Control Tree Recognize flagged control results for COULTER 5C Cell Control and COULTER Retic-C Cell Control Access HELP information for action to take when a control is outside expected ranges Deselect 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 the most accurate results possible to the clinician. Knowing how to review and output control results is an important follow-up to control processing. It allows recognition of shifts or trends in QC that might indicate control or instrument problems. Resources To complete this module you will need LH 500 LH Workstation with HELP Instructions for Use (IFU) for LATRON PRIMER and CONTROL, 5C Cell Control, Retic-C Cell Control Control products for processing Retic Prep kit (Optional) Control folders with stored data B-37 of 68

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49 INFORMATION / PRACTICE SESSION Control Storage and Handling Refer to the Instructions for Use 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. Get the 5C and Retic-C control vials for your unit from the refrigerator to begin equilibrating at AMBIENT TEMPERATURE (per Instructions for Use) as you do these next steps. B-39 of 68

50 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) NOTE: If you have never used LATRON Primer and Control, see a facilitator for additional details about using these products. Step 1 Action Run the Latron Primer and Control found at your unit following the Running Latex Control --Diff and Retic procedure from HELP. Watch out for these relatively common errors! Tips 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. 2 Use the Control tree to view your Latron results. 3 4 Note that you have two entries with the same date, one for the Primer and one for the Control. Use both horizontal scroll bars to view all results for both 5PD (fivepart diff) and Retic. Control Primer Primer Scroll bar Control Scroll bar B-40 of 68

51 Latex Control Is Outside Expected Ranges COULTER LH 500 TRAINING MODULES Step Action At the Workstation go to on the Command Center. Choose on the Common Toolbar. And then from the Common Toolbar. Topic on the screen is: Reviewing Control Results In the HELP window at the left of the screen, select the Contents tab, then LH 500 Analyzer, then Controls. Choose When a Latex Control is Outside its Expected Ranges. Choose display. Read the topic. You now have some basic troubleshooting tools to use if LATRON Control is outside its expected ranges. 4 Close HELP using the in the upper right corner. Running 5C and Retic-C Cell Controls Step 1 Action Open HELP from the Common Toolbar, select Index and type CYCLING, click on and Display Cycling Controls in the Automatic Aspiration Mode. 2 Read the procedure. 3 Close HELP. NOTE 4 5 You do not have to be on the QA/QC screen when you run controls Use the HELP procedure to make the proper setup selections on the Command Center. Place your properly mixed controls into cassette(s) with bar code labels facing up. 6 Place cassette(s) into the right-hand loading bay. 7 8 Use the Control Tree (See the Control Tree on the next page) to access control folders and ensure that control results are in their correct folders. Return the 5C Cell Control and Retic-C Cell Control vials to the refrigerator within 30 minutes. B-41 of 68

52 LH 500 Control Tree ABN II ABN I Norm Level I Level II Level III 5C and/or Retic-C Cell Controls are Outside the Expected Ranges Step Action 1 At the Workstation QA/QC screen select (from the Common Toolbar). 2 Use HELP to find the following topic: When a Control is Outside its Expected Ranges 3 Print 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. B-42 of 68

53 Finding Flagged Control Results Step 1 Action 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 order. in front of 5C. All 5C folders open in numerical 3 4 NOTE Click directly on the 5C Cell Control folder you wish to open. The most recent result is at the top of the list. Search through the 5C folders 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. If you cannot find a control run that is out of limits, then choose any control run in any control folder (including Latron). Deselecting and Restoring a Control Result Step Action 1 Now that you ve found a flagged result, click on the deselect [Des] button at the left of the line. The line is now grayed out and removed from the statistical results below. Note the total number of runs in the control folder after removing one run. NOTE When you print a file with a deselected run, the word yes appears in the deselect (des) column. 2 Practice deselecting and restoring control results, but leave at least one removed result in the folder for your final printout. B-43 of 68

54 Add a Comment in a Control Folder Step Action 1 Choose a control run from today. Click on the button. 2 Type a comment in the comment box and select comment. [OK] to save the 3 4 NOTE 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. Click on any box with an X in the CMNT column to view a comment, add text or change text. When you print the file, the comment appears under the run with which it is associated. 5 Select (History Logs) from the Command Center. 6 Select the Control Data Tab and view the comment added to the control file. 7 Close the History Logs using the in the upper right corner. Levey-Jennings Graphs Step 1 Action Open the QA/QC window again. Select the same control folder, which you added a comment, to work with. Below the statistical information for a control folder are the thumbnail Levey-Jennings graphs showing the last 10 runs in the control folder. 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 the graphs for other parameters. 2 3 Scroll to PLT and click on it now. Use the graphs to observe trends and shifts. Double-click on any thumbnail graph to see a larger version called full-page. This graph shows up to 100 data points for the selected control folder. Do this now. 4 Close the full-page graph by selecting Exit. B-44 of 68

55 Printing Control Results Step COULTER LH 500 TRAINING MODULES Action 1 With the control folder still open, from the Common Tool Bar select the (Print) button. By selecting the desired radio buttons on this screen you can print various combinations of control folder data. For this exercise, choose Selected Lot and Thumbnail graphs. 2 3 After making your selection, click on. 4 Collect your printouts and save for the Skill Check. 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 control runs by shift. Shift 0 is all runs from all shifts. Shifts 1, 2 or 3 are the runs by individual shift as definded by your lab. 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. To set up individual shift times go to: System Setup Quality Assurance. B-45 of 68

56 Practice Step Action Select System Setup and Select the Select 3 tab. Use Multiple Shifts View the area where shift times are entered. Please do not change. View what is setup on your unit. Press (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 above. 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 at 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, proceed to the Skill Check for this module. Collect your printouts and proceed to the Skill Check for this module. B-46 of 68

57 QC COULTER LH 500 TRAINING MODULES 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? (4 answers) B-47 of 68

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59 RPS COULTER LH 500 TRAINING MODULES 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) B-49 of 68

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61 INFORMATION / PRACTICE SECTION Sample Storage and Stability Step 1 2 Action At a Workstation, from (HELP), go to Contents LH 500 Analyzer Operating Operating-BASIC Display and print the following topics Collecting Specimens Storing Specimens. 3 Close HELP 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? 4 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 Be sure to wear your labcoat and gloves, during this procedure. Before you begin, make sure you have the supplies you need: Blood samples collected in EDTA Bar code labels Sample cassettes B-51 of 68

62 Instrument/Workstation Overview Step Action 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. Click on (Run Configuration) from the Command Center. Make sure that Automatic Output, Print (tab) is set to All Samples At the Command Center, select AUTO ANALYSIS as the Process Type, CD as the Run Type and AUTO as the Asp Mode. Select Start. Running Samples Step Action Place CD sample tubes into a sample cassette, making sure the bar code labels are facing up. 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. 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. B-52 of 68

63 Manual Mode COULTER LH 500 TRAINING MODULES 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. Step 1 Action 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 NOTE 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. Make sure the sample is well mixed. Remove the stopper from the tube. 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. 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! B-53 of 68

64 Running a STAT Step 1 Action 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 6 Run the Stat in manual mode. When the cycle completes, select stop. 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. Step Action 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. On the Workstation, Command Center: 2 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 B-54 of 68

65 NOTE 3 4 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. Identify your sample to the LH 500 using the keyboard to type an ID. Press Enter or Tab. 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. Using Micro-collection Tubes (Elective Exercise) Step Action Get micro-collection tubes, EDTA blood tubes, and transfer pipettes from the supply in the lab. 1 2 Look at a demo micro-collection tube for the fill level required for manual mode aspiration. 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. 6 Process sample in the manual mode. 7 For additional samples, repeat steps Save printouts for the Skill Check. B-55 of 68

66 Running Samples in the Retic Mode (Elective Exercise) Step Action 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. 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 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. 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. 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. B-56 of 68

67 Using Navigation Buttons COULTER LH 500 TRAINING MODULES 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. 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: 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. 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. B-57 of 68

68 Adding a Sample Request to the ToDo List Step Action 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. B-58 of 68

69 RPS COULTER LH 500 TRAINING MODULES RUNNING PATIENT SAMPLES SKILL CHECK Have your facilitator check your answers to the questions on page B-51 of 68 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? B-59 of 68

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71 SD COULTER LH 500 TRAINING MODULES 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 B-61 of 68

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73 INFORMATION / PRACTICE SECTION 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 4:15. Step 1 Action From the Command Center select Stop SELECT FUNCTION. to put Instrument Status to 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. B-63 of 68

74 7 NOTE If you need to interrupt the Clean cycle you have two choices F4: Abort the cycle F5: Run Startup now It is preferable to use F5 if you cannot wait the entire 30 minutes (e.g. a stat request) Clean Cycle timing is NOT programmable or changeable, other than interrupting if necessary, as indicated above. Shutdown Step 1 Action From the Command Center select Stop SELECT FUNCTION. to put Instrument Status to 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 B-64 of 68 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: o Brings up the compressor o Purges the flow cell and sample line with diluent o 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.

75 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 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 B-65 of 68

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77 SD COULTER LH 500 TRAINING MODULES 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? B-67 of 68

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79 SETUP MODULES Implementation Troubleshooting Setup Basics This document is not intended to replace the information in your Instrument Instructions for Use Manual (IFU). Information in the Instructions for Use Manual supersedes information in any other manual. S-1 of 40

80 You will see the following symbols used throughout the learning modules. Important information you should know. Techie-type information you may be interested in knowing. Do these steps now! Timely Tip You reached the goal! I do not understand Questions Resources, Help S-2 of 40

81 COURSE MAP - SETUP Begin Here Reagent Setup and Replacement Setting Up Controls 1 Additional QC Calibration S-3 of 40

82 Name PERSONAL PROGRESS SUMMARY SetUp Modules Date Initial Facilitator Reagent Setup and Replacement Setting Up Controls 1 Additional QC Calibration S-4 of 40

83 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 S-5 of 40

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85 INFORMATION / PRACTICE SECTION COULTER LH 500 TRAINING MODULES 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 reagents. The open expiration is the date to which an opened container may be used, based on the Date Opened. S-7 of 40

86 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. S-8 of 40

87 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. S-9 of 40

88 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. S-10 of 40

89 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. S-11 of 40

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91 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. S-13 of 40

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93 SC1 COULTER LH 500 TRAINING MODULES SETTING UP CONTROLS 1 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 assay sheet Set up COULTER 5C Cell Control and COULTER Retic-C Cell Control folders 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 assay sheet 5C Cell Control assay sheet with 2D barcodes Removable Media (CD, Flash drive) S-15 of 40

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95 INFORMATION / PRACTICE SECTION COULTER LH 500 TRAINING MODULES Step Action From the Command Center go to [System Set Up] 1 [Quality Assurance Set Up] (tab) Line Select Set up Lab Limits Set up New Control Folder Archive Edit values Transmit Delete selected files 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. 2 Select [New Control Folder] 3 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 4 Select [Set Up New Lot] S-17 of 40

96 5 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. 6 Select [OK] 7 When you are setting up LATRON, after selecting the button, you enter information from the package insert starting with Lot # and expiration date. Warning 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. 8 Select [OK]. 9 Select [Close] to close the Setup New Control Folder window. 10 Select window. [OK] to close the System Setup / Quality Assurance 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. S-18 of 40

97 SC1 COULTER LH 500 TRAINING MODULES SETTING UP CONTROLS 1 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. S-19 of 40

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99 QC2 COULTER LH 500 TRAINING MODULES 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 S-21 of 40

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101 INFORMATION / PRACTICE SECTION IQAP Download Procedure Step 1 Action In HELP go to Index 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. 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. S-23 of 40

102 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. Other IQAP Considerations 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. S-24 of 40

103 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. S-25 of 40

104 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. S-26 of 40

105 QC2 COULTER LH 500 TRAINING MODULES 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? S-27 of 40

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107 CAL COULTER LH 500 TRAINING MODULES CALIBRATION OBJECTIVES Given an operating COULTER LH 500 and access to HELP Perform calibration of an LH 500 Analyzer using COULTER S-CAL Calibration Kit Adjust calibration factors as required by the calibration results Identify when you should calibrate your instrument Identify when you should verify calibration of your instrument Why Is It Important? Calibration fine-tunes the LH 500 to give the most accurate results possible. Resources To complete this module you will need LH 500 Analyzer LH Workstation with HELP One vial of COULTER S-CAL from the Calibration Kit One tube of EDTA blood S-CAL assay sheet (included in S-CAL package) with 2D barcode S-CAL Instructions for Use (see facilitator for a copy) 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. S-29 of 40

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109 INFORMATION / PRACTICE SECTION 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 components involving dilution characteristics or primary measurements need to be replaced. After the replacement of any component that involves dilution characteristics (such as a BSV) or primary measurement (such as an aperture), your service engineer will perform a rough calibration afterward using your current controls to allow instrument operation to continue. Keep in mind that a new component should be given several days to break-in or settle before performing the verification. An S-CAL verification should be performed within several days to a week to verify cal factors. Calibrating CBC Parameters with S-CAL Calibrator Use the on-line HELP system to find the topic, Calibrating CBC Parameters with S-CAL Calibrator. You will see that there are nine main steps to complete calibration on the instrument. Note that each of the nine steps are linked via hypertext to the details for that step. 1. Ensure the Apertures are Clean If you routinely shut down the instrument for at least 30 minutes every 24 hours in COULTER Clenz, and the instrument is currently in Shutdown, cleaning is unnecessary. If you routinely use COULTER Clenz, but you are beginning calibration after processing patient samples, perform a shutdown for 30 minutes before proceeding. You could use the Clean Cycle if you wish. (You will not need to do this in the classroom.) If you do not routinely use COULTER Clenz, then you must bleach the apertures before proceeding with calibration. Follow the HELP link if this is the case. (You will not need to do this in the classroom.) CAUTION Possible flow cell damage could occur if you aspirate bleach. Do Not Aspirate Bleach. S-31 of 40

110 2. Verify Instrument Functions Properly Note: Both the Reproducibility and Carryover procedures may be a requirement of regulatory agencies. It is no longer required to perform these procedures prior to calibration; however, it is good lab practice to do so. Use these procedures at any time to verify instrument performance. Refer to the Performance Specifications in HELP for the results criteria. Step 1 2 Action Check the reagent containers for: Sufficient quantity For your lab: diluent container approximately 1/2 to 2/3 full; CBC lytic reagent at least ¼ full 1L or 5L Check with an instructor before changing any classroom reagents Not beyond expiration date No precipitates, turbidity, particulate matter, or unusual color Proper connections between the Diluter and the reagent containers Check the waste container (if present) for: Sufficient capacity Proper connections 3 Perform Startup. (This is not necessary for class purposes, if Startup has already been performed.) 4 For class purposes: (not necessary if already run) Run COULTER LATRON Primer and Control and COULTER 5C Cell Control. Confirm that results are within limits. S-32 of 40

111 3. Prepare Instrument for Calibration COULTER LH 500 TRAINING MODULES 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). Step 1 Action Ask an instructor for a vial of S-CAL. Allow it to warm up at room temperature for 15 minutes before using. (See Instructions for Use for details. IFUs can be found on the BCI website.) 2 On the Workstation Command Center, ensure the blood detector is enabled (checked). 3 Set the Process Type to Auto Analysis. 4 Set the Run Type to C 5 Set the number of aspirations per tube to 1. 6 Select Start 7 Cycle a sample of normal whole blood in automatic aspiration mode as a prime. 8 Select Stop 9 Change the number of aspirations per tube to 11. S-33 of 40

112 4. Calibration Set Up at the Workstation NOTE: If your instrument does not have a 2D-barcode scanner, please refer to the next section on Downloading Using Removable Media (next page). In class we will use the 2D-barcode scanner. Step ACTION 1 At the INSTRUMENT Workstation Command Center, select CALIBRATION in the Process Type drop-down box. 2 Go to on the Command Center then select (CBC Calibration) on the Common Toolbar. 3 Note The background color of the Calibration window changes to the same background color as the Command Center when you change Process Type to CALIBRATION. Forgetting to do this is a very common error. If you forget to change the process type to CALIBRATION, all S-CAL runs will be analyzed as patient samples and there will be NO data in the calibration table. 4 If necessary, select (Clear table) to clear old calibration runs from the database. The values on the table change to 0.00 except for Old Cal Factor. 5 On the specific toolbar, select (Calibration Setup) to display the Instrument Calibration Setup window. 6 Select (Load S-Cal values). Scan the 2D-barcode for your instrument (LH 500) on the S-CAL assay sheet to load the reference values, lot number and expiration date. Ensure that you select the assay values for the reagents on your instrument. 7 Print the Instrument Calibration Setup window. Mark it precal. Save it for the Skill Check. 8 Select on the Instrument Calibration Setup window. 9 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. S-34 of 40

113 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 backlit in RED with the message Calibrator expired. No statistics will be calculated. appears in the task bar. 5. 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 Instructions for Use. Step 1 Action Prepare the S-CAL calibrator vial according to the Instructions for Use. (Two sets of 8x8x8, just like the 5C Cell Control). It is very important to follow the mixing directions carefully. 2 Verify the number of aspirations is 11. (One vial of S-CAL is sufficient for 11 aspirations.) 3 At this point, ensure that the blood dectectors are enabled, CALIBRATION is chosen in the Process Type box on the Command Center and the current S-CAL lot # is in the drop down box on the calibration table window. 4 Place the S-CAL vial into a cassette, and place the cassette in the right loading bay on the Diluter. 5 6 Select Start Automatic processing of the cassette begins. The Workstation automatically deselects the results from the first run because those results are used as a prime. S-35 of 40

114 6. Review Results On The Calibration Window 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 selected parameter, deselect the check box. 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 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. 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 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). If you choose to transmit calibration factors other than those selected by the Workstation either outside 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. S-36 of 40

115 CALIBRATION CRITERIA TABLE (This table is provided for your information only. The workstation makes these calculations and decisions for you). 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 0.8% 2.0% >0.7% BUT 2.0% >0.03 BUT 0.09 Hgb CV 0.8% 3.0% >0.78% BUT 3.0% >0.1 BUT 0.4 MCV CV 0.8% 2.5% >1.18% BUT 2.5% >1.0 BUT 2.0 Plt CV 3.2% 9.0% >2.70% BUT 9.0% >6.0 BUT 20.0 MPV CV 5.0% 20.0% >5.0% BUT 20.0% >0.5 BUT 2.0 Are all parameters within limits? Are any boxes checked? If No, then no further action is necessary. Calibration is verified. 7. Adjust Calibration Factors As Needed Step Action 1 Are any boxes checked? If Yes, then parameter(s) need to be calibrated. If No, contact the facilitator before proceeding. 2 On the Workstation CALIBRATION screen, select (Adjust Calibration) on the Specific Toolbar. 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. 4 Select to print the calibration runs for your logbook in your lab. Save it for the Skill Check in class. Don t do this next step until the skill check, when the facilitator is present. 5 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. 6 Select (Calibration Setup) to display the Instrument Calibration Setup window and reprint this window. Mark it post-cal and save for your Skill Check. S-37 of 40

116 7 On the Command Center, set Process Type back to Auto Analysis and at the #aspirations/tube back to ONE. 8 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. If you run S-CAL calibrator and change at least one parameter to a new cal factor, you have calibrated that parameter. You 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. Calibrate or Verify Calibration? Step Action 1 From the CALIBRATION screen, select on the Common Toolbar. 2 Click on the Index tab. 3 Type calibration. 4 From the list on the left, select the topic: Calibration Overview. 5 Review the information about When to Calibrate to discuss as part of the Skill Check for this module. Now that you have completed the calibration process, take a look at the summarized procedure in the Easy Reference Guide. This provides a short summary of the steps of calibration without all the details. You may use this summary in your lab. Remember that you may always refer back to the online HELP procedure or to this module for the details. Proceed to the Skill Check for this module and keep the following with you: Calibration Set Up (one before running S-CAL (pre-cal) and one after transmitting new Cal factors (post-cal)) and Calibration Run screen printouts (three total) Calibration overview printout S-38 of 40

117 CAL 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 Table and two Calibration Set Up screens- pre-cal and post-cal ) 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. Choose all of the valid reasons to run S-CAL calibrator to verify calibration settings? (See Calibration Overview in help.) 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 Have the facilitator show you how to and why to clear the calibration table. S-39 of 40

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119 TROUBLESHOOTING MODULES Implementation Troubleshooting Setup Basics This document is not intended to replace the information in your Instrument Instructions for Use Manual (IFU). Information in the Instructions for Use Manual supersedes information in any other manual. PN CA.1 (June 2012) TB-1 of 74

120 You will see the following symbols used throughout the learning modules. Important information you should know. Techie-type information you may be interested in knowing. Do these steps now! Timely Tip You reached the goal! I do not understand Questions Resources, Help TB-2 of 74 PN CA.1 (June 2012)

121 COURSE MAP - TROUBLESHOOTING Begin Here History Logs Sample Flow Replacing Components Troubleshooting Basics TB-3 of 74

122 Name PERSONAL PROGRESS SUMMARY Troubleshooting Modules Date Initial Facilitator History Logs Sample Flow Replacing Components Troubleshooting Basics TB-4 of 74 PN CA.1 (June 2012)

123 HL COULTER LH 500 TRAINING MODULES 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 Access detailed descriptions and corrective actions for messages Print a History Log Archive a History Log (optional) 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 CD or Flash Drive TB-5 of 74

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125 INFORMATION / PRACTICE SECTION COULTER LH 500 TRAINING MODULES Step Action 1 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. 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 Read the topic, Working with Electronic History Logbooks, as well as the data from the add comments to a logbook (hyperlink) 4 Close HELP. Practice Use the LH Workstation at your instrument for the Practice part of this module. Adding a Comment Step Action 1 Select any History Log, using the tab. Which tab did you choose? 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. TB-7 of 74

126 Printing from a History Log Step Action 1 Select the Calibration History Log. 2 Select the Print button. 3 Choose Selected Category. 4 Choose By Date:. 5 Enter the date two months ago in the From box and today s date in the To box. 6 Select. 7 Save the printout for the Skill Check. Deleting Note It is not necessary to delete to make room in the history log. Each tab in the history log holds 2500 entries, then it rolls. The oldest entry is deleted and the most recent is added (first in, first out). It is not recommended to delete any entries in the history log as the information in this section may help when troubleshooting. TB-8 of 74 PN CA.1 (June 2012)

127 Archiving (Optional Exercise) Archive Using Removable Media (CD-ROM or Flash Drive) Step Action 1 Select any History Log. 2 Select the button. [Archive Result(s)] 3 Choose the Selected Category radio button. You can specify by date if you wish. Click on the OK button. 4 A Save As box appears with the message A:/ is not accessible. The folder was moved or removed. Click on Okay. 5 Another Save As box appears. Place your removable media into the appropiate drive. 6 In the Save As box, specify the drive where you want to archive the information. 7 Type in the file name or use the suggested name which is the current date and then click on Save. Close the History Logs Viewer window If using using a CD-ROM, press the eject button on the CD drive on the Workstation computer. A Drag-to-Disc dialog box with the message Failed to Rename disc appears. Choose OK. A Drag-to- Disc Eject Options dialog box opens. Select This disc will be used on other computers or devices. Keep enabled the Always show this dialog when ejecting a disc. Then choose the Eject button. Another Drag-to-Disc box with a progress bar opens. Wait for it to become 100% complete. The CD drive will eject the CD. Save your archive for the skill check. If you can perform all the operations from the Practice part of the module, collect your data and printouts and proceed to the Skill Check. TB-9 of 74

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129 HL COULTER LH 500 TRAINING MODULES HISTORY LOGS SKILL CHECK Collect the media you used for archiving, as well as the required printouts. 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. TB-11 of 74

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131 SF COULTER LH 500 TRAINING MODULES 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 TB-39 of 74. TB-13 of 74

132 SET UP When operating the LH 500 Analyzer in your laboratory you must follow 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. Step Action 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 2. 2 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. Ask facilitator to place the magent. 5 Open the right side door pulling from the left. 6 For cycling the LH 500 Analyzer, use the tube provided by a facilitator. TB-14 of 74 PN CA.1 (June 2012)

133 INFORMATION / PRACTICE SECTION Module and Component Identification 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. TB-15 of 74

134 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 crews Needle S h ield TB-16 of 74 PN CA.1 (June 2012)

135 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 TB-17 of 74

136 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 TB-18 of 74 PN CA.1 (June 2012)

137 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). TB-19 of 74

138 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. TB-20 of 74 PN CA.1 (June 2012)

139 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 TB-21 of 74

140 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. TB-22 of 74 PN CA.1 (June 2012)

141 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 TB-39 of 74. TB-23 of 74

142 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 TB-24 of 74 PN CA.1 (June 2012)

143 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. TB-25 of 74

144 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? TB-26 of 74 PN CA.1 (June 2012)

145 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 TB-27 of 74

146 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 TB-28 of 74 PN CA.1 (June 2012)

147 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. TB-29 of 74

148 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 Principle, refer to the Hematology Innovations prerequisite, 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) TB-30 of 74 PN CA.1 (June 2012)

149 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. Go to the Component Table in the lab to look at an aperture bath. Remove an aperture block to see the aperture. 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 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 TB-31 of 74

150 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. TB-32 of 74 PN CA.1 (June 2012)

151 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. Go to the Component Table in the lab to see the coaxial cables connected to the back of the aperture bath. 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 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. TB-33 of 74

152 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. TB-34 of 74 PN CA.1 (June 2012)

153 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. TB-35 of 74

154 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. TB-36 of 74 PN CA.1 (June 2012)

155 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. TB-37 of 74

156 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. TB-38 of 74 PN CA.1 (June 2012)

157 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. TB-39 of 74

158 TB-40 of 74 PN CA.1 (June 2012)

159 SF COULTER LH 500 TRAINING MODULES 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 Analyzer Component Checklist 1. Rocker bed 13. CBC lytic reagent pumps 2. Tube ram 14. WBC diluent dispenser 3. Tube available sensor 15. RBC Bath mixing bubble check valve 4. Piercing needle cartidge 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 TB-41 of 74

160 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) Principle List the four constants necessary for proper application of the Principle when counting and sizing particles: Name the variable in the application of the Principle Sample Flow Write an explanation of the importance in the diluter cycle for each of the following. Sweepflow Mixing bubbles Cross-rinse TB-42 of 74 PN CA.1 (June 2012)

161 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. TB-43 of 74

162 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. TB-44 of 74 PN CA.1 (June 2012)

163 INFORMATION / PRACTICE SECTION TUBING Types of Tubing I-beam Round 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 Pharmed 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. Almost always used in pinch valves through the normally open part of a pinch valve. Color-code stripe Blue, Black, Yellow, Red, Green, Brown. 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. TB-45 of 74

164 PINCH 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. ANGAR 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 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. TB-46 of 74 PN CA.1 (June 2012)

165 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 TB-47 of 74

166 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 TB-48 of 74 PN CA.1 (June 2012)

167 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 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. TB-49 of 74

168 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. TB-50 of 74 PN CA.1 (June 2012)

169 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. TB-51 of 74