BAUSCH & LOMB ENGINEERING ROCHESTER. Title: Membrane Press Procedure Page 1 of 14

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Title: Membrane Press Procedure Page 1 of 14 Author: K. Schober Attachments: 3 Released by: Date: 5/5/06 Global Document Management DCO-SOPROC PURPOSE/SCOPE To provide instructions for how to conduct the test of the biological tissue (rhexis). RESPONSIBILITY It is the responsibility of all the operators to read through this procedure prior to beginning any testing, and to follow this procedure to correctly and safely perform the necessary testing. MATERIALS AND EQUIPMENT Equipment Linear Stage (S/N B06 0049) Load Cell (94062) Forceps (Peer-Vigor 57-413) Controller (ESP100) Saline Bath (11304) Sample Mount (11301/11302) Amplifier Materials Wipe (TechniCloth TX606) Isopropyl Alcohol 99% (IPA) RM 1290 O-ring (S70-M.38X3.37, S70-M1X2) MEMBRANE PRESS SETUP This procedure demonstrates how to setup the device prior to testing. Step Action 1 Plug in the Newport Controller and the USB connector or RS232 cable, whichever is compatible with the computer. Turn the power on.

Title: Membrane Press Procedure Page 2 of 14 2 Plug in the Amplifier. 3 Plug the USB from the Data Acquisition card into the computer.

Title: Membrane Press Procedure Page 3 of 14 4 Open the ESP-Util Program. A Select Communications Port pop-up should be up. Select the Serial (RS232) as the port type. Select preferred Com Port. Click Open Port. Click Yes to Reset Controller. After all this has been completed, you may close out of the ESP-Util Program (Either click the X in the top-right corner or File-Exit). 5 Open Labview Programs named, AlignmentVI_revised on 4-28-06 and Complete Test Procedure. 6 Gently screw probe tip attachment into the load cell. NOTE: Load cell pins MUST be in the load cell to screw the probe tip in. Probe tip attachment Load Cell pins

Title: Membrane Press Procedure Page 4 of 14 SAMPLE PREPARATION This procedure demonstrates how to prepare the sample and load sample mount prior to testing. Step Action 1 Using forceps take an o-ring (red) and place in designated location on mount. Should be a snug fit. Bottommount piece. O-ring 2 Using the forceps take the material and carefully place it over the o-ring. Make sure that are no creases and that the material is resting flat over the opening with no sagging. NOTE: To make this step easier you could use a microscope while placing the material over the o-ring.

Title: Membrane Press Procedure Page 5 of 14 3 Place the top mount piece on the bottom mount. Use the three pins protruding from the bottom mount to guide the top. Bottom-side of top piece 3 pins on bottom piece NOTE: The top-side of the top piece looks like this: Notice that the hole looks larger.

Title: Membrane Press Procedure Page 6 of 14 4 Pull and stretch the 3 knobs up and over the top piece. Make sure that they rest in their notch. Before After 5 1. If testing material in saline solution, skip to step 7 2. If testing dry, proceed with step 6. 6 Place mount into saline bath. Saline bath 7 Off to the side fill a large container with saline solution. Skip to Membrane Press Alignment procedure. 8 Taking the saline bath piece and the sample mount piece slowly submerge both separately into the container. Angling the two pieces while inserting the sample mount into the saline bath piece. NOTE: The key to this is eliminating as many air bubbles as possible.

Title: Membrane Press Procedure Page 7 of 14 MEMBRANE PRESS ALIGNMENT This procedure demonstrates how to align the probe tip to the center of the micrometer stage. This procedure must be completed after each test. Step Action NOTE *The micrometer slides should have been in the locked position prior to alignment. If the knobs weren t locked BE CAUTIOUS when lowering the probe. 1 Carefully place sample mount (from Sample Mount Preparation) into its designated location watching out for the probe tip. Sample mount rests in here. 2 Carefully remove the two pins from the load cell.

Title: Membrane Press Procedure Page 8 of 14 3 Use the AlignmentVI_revised on 4-28-06 LabVIEW program to lower stage. Click the white arrow in the top left corner, and then click the Reset Motor back ON button. White arrow Forc e dial This is where you type in desired position. 4 Lower the stage to position 19. Type, 19.000 into the Target Position, then click Move. The probe tip will not hit the mount at this position. 5 Closely look at the probe tip and the hole to verify that the probe tip will not come in contact. 6 Lower the stage to position 20 (using the same method in Step 4), while checking the force dial on the computer screen. After the stage has stopped it should read noise from the load cell. Ranging from -.00892g to.00892g. NOTE: There is an overload built into this program, if the load cell reads a force of 5 grams or greater the stage will immediately stop. If this occurs raise stage to position 0 using step 4. 7 Repeat Step 4, lowering the stage to position 21. Looking at the hole and the tip. Use the micrometer stage if adjustments need to be taken. At this position the tip will be half way in the whole. 8 Repeat Step 4, lower stage to 22. This is the position you ll align the probe and stage following the next steps listed out. NOTE: At this position the range of force should be -.00892 to.00892, which is noise.

Title: Membrane Press Procedure Page 9 of 14 9 Using the x-direction micrometer slide. Move the stage to the left until the force dial reads +/- 0.05352g. Read and record the position on the slide on appropriate data sheet (See Attachment 1). Turn knob in this direction will move stage to the LEFT. LOCKS stage Turn knob in this direction will move stage to the RIGHT. 10 Repeat Step 10, moving stage to the right side. 11 Take the measurements from Step 10 and 11; take the average of the two. Move the x-direction slide to the average location. NOTE: The slide will need to be moved to the left. 12 Using the y-direction micrometer slide. Move the stage to the bottom until the force dial reads +/- 0.05352g. Read and record the position on the slide on appropriate data sheet (See Attachment 1). LOCKS stage Turn knob in this direction will move stage to the LEFT. Turn knob in this direction will move stage to the LEFT. 13 Repeat Step 13, moving stage to the top. 14 Lock stage in y-direction. 15 Repeat Steps 10-12. 16 Lock stage in x-direction.

Title: Membrane Press Procedure Page 10 of 14 TEST EXECUTION AND DATA ANALYSIS This procedure demonstrates how to run a test and analyze the data. Step Action 1 The AlignmentVI_revised on 4-28-06 LabVIEW program should still be up from the alignment procedure. Click the Red Stop Sign button in the top left corner. 2 Open the Complete Test Procedure LabVIEW program. Click the white arrow and then click the Move button. This button initiates the test. NOTE: If for any reason the test needs to be terminated, click the Emergency OFF! button. 3 The test will be completed if the overload is reached (5 grams) or if the stage reached its max position (25). When the test is complete the stage will automatically raise itself. 4 A window will pop-up and ask for a location for the results. Save in designated location. When typing in the file name place a.txt after the name. This will make the data a text file, which will be needed for data analysis. 5 Click the Red stop sign button in the top left corner of the labview program. 6 If finished running tests, following the test carefully place the load cell pins into the load cell. If not finished you may leave pins out but be cautious.

Title: Membrane Press Procedure Page 11 of 14 7 Open the Data Analysis Excel spreadsheet. 8 Open the text file you previously saved. Click Ctrl+A and then Ctrl+C 9 Paste this data into the excel spreadsheet. 10 Click Process Data button on the excel spreadsheet. 11 Record the Young s Modulus value and R 2 into Attachment 2. 12 When closing out Excel, click No to save changes. Do the same for the LabVIEW programs. HISTORY Revision A: DCR- Original.

Title: Membrane Press Procedure Page 12 of 14 Attachment 1: Alignment Table Operator: Date: X-Direction Y-Direction After Test # Left side Right side Center After Test # Bottom Top Center 0 0 0 1 1 2 1 3 2 4 2 5 3 6 3 7 4 8 4 9 5 10 5 6 6 7 7 8 8 9 9 10 10

Title: Membrane Press Procedure Page 13 of 14 Attachment 2: Young s Modulus Table Operator: Date: Test # Young's Modulus (N/mm 2 ) R 2 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Title: Membrane Press Procedure Page 14 of 14 Attachment 3: Packing the device into the case.