Sample Preparation and Setup of BioPress ™ Culture Plates and
Contents
1. Tech Report 111 Sample Preparation and Setup of BioPress Culture Plates and Compression Baseplate Assembly Author Chris Wimmer Document BioPress and Compression Baseplate Assembly Tech Report Rev 1 1 01 07 2015 Dunn Labortechnik GmbH Thelenberg 6 53567 Asbach Tel 49 0 26 83 4 30 94 Fax 49 0 26 83 4 27 76 e mail info dunnlab de Internet www dunnlab de COPYRIGHT 2009 FLEXCELL INTERNATIONAL CORPORATION FLEXCELL INTERNATIONAL CORPORATION The BioPress compression culture plate contains a foam ring and sample holder in each well Fig 1 The foam ring is used to contain the removable sample holder In preparing samples be sure that the sample thickness is no less than 1000 um and no greater than 0 118 3 mm The thickness of the compressed foam is 350 um therefore thinner samples will not be compressed A sample thickness of 1000 um or more will ensure compressibility The maximum chamber height is 0 122 3 1 mm therefore samples with a thickness greater than 0 118 3 mm will be preloaded when the platens are inserted into the wells of the culture plates Figure L Compression culture plate foam ring and sample holder The sample diameter should be equal to or less than the inner diameter of the foam sample holder in the chamber which is 0 200 5 mm If the user wishes to use a larger sample they can remove the sample holder and place the sample withi2. ight in inches x 0 122 h 0 025 where x is the of 180 degree clockwise turns required for the center screw to touch the sample and h is sample height in inches NOTE If the sample height is larger than 3 11 mm 0 122 x in the above equations will be a negative number In this case the bottom of the center screw will compress the sample when it is flush with the bottom of the platen body as the platen is first inserted into the well To compensate for this the center screw can be turned counterclockwise x number of turns however it should be noted that the sample will no longer be fully compressible as the base of the sample holder will no longer be able to contact the base of the center screw Using sterile forceps turn the center screw clockwise the number of turns required by the above equation s Be careful not to screw the center down too far as this will preload the sample and give inaccurate force readings Add or remove fluid medium through any of the eight holes around the inner periphery of the platen Fig 2 Center the BioPress compression plates on top of the four black gaskets in the baseplate Fig 3 Adjust the eight clamping pads in the clamping system to their maximum height and turn them so that the bottom of each pad is parallel with the surface of the baseplate Place the Plexiglas window into the clamping system Fig 4 evenly along the bottom four clamping bars NOTE The Plex
3. iglas window is used as a backing support for the baseplate to enable it to withstand high pressures Never operate the compression baseplate without this window in place Baseplate without plates Baseplate with plates Figure 4 Clamping system with Plexiglas window E FLEXCELL INTERNATIONAL CORPORATION Carefully slide the baseplate plate assembly into the clamping system so that the four BioPress plates are centered underneath the eight clamping pads Fig 5 Check the Plexiglas window and move as necessary to center it under the baseplate Turn the wing nuts on the clamping pads until they are finger tight This will compress the plates downward onto the gaskets creating a seal Baseplate slides into into system Plexiglas indow on bottom and then t hten the clamps down over he compression plates Do Plexiglas window must fit Tinder the baseplate to feep seal Figure 5 Clamping system with baseplate and culture plates NOTE If using less than 4 plates in an experiment use an old plate or support in place of the missing plate s and tighten the clamping pads onto this surface This will ensure that the baseplate is seeing an even and sufficient pressure across its surface and will prevent baseplate blowout Place the entire assembly into the incubator Attach the FLEX IN and FLEX OUT quick disconnect fittings to the fittings on the com
4. n the larger ring that normally contains the sample holder This will allow a sample diameter of up to 0 525 13 mm To prepare a sample for loading remove the sample holder from the plate using a pair of sterile forceps Place the tissue or gel sample into the hole at the center of the removable sample holder Carefully place the sample holder back into the BioPress compression plate so that it is contained within the foam ring Do this for the remainder of the wells Add 3 5 mL of culture medium to each well Prepare all stationary platens by adjusting the center screw Fig 2 such that the screw bottom part that touches the sample is exactly flush with the bottom of the platen body Use a pair of sterile forceps and the two holes at the adjustable center of the platen to turn the center screw j f Hole for fluid J medium additior d iustable cent Figure 2 Stationary platen and compression culture plate Place a stationary platen into each well of the compression plate Fig 2 Calculate the number of revolutions required to bring the bottom of the center screw to the point where it just touches the top of the sample by using the following equations For sample height in millimeters x 3 11 h 0 62 where x is the of 180 degree clockwise turns required for the center screw to touch the sample and h is the sample height in millimeters FLEXCELL INTERNATIONAL CORPORATION For sample he
5. pression baseplate Samples are now ready for compression loading Please see the Compression User Manual for further instructions on running a compression regimen
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