i seeded CACO2 on 0,4 um PET transwell insert, i want to see my cell layer in transversal view in order to monitor the polarization and formation of brush border. To do this, i fixed with 4% PFA in PBS for 30 minutes, then put in alchol 70% until the processing.
I continued the dehydratation with 95% alchol for 1h (twice), 100% alchol for 1h (three times), xylene for 1h (twice), paraffine overnight, and one change of paraffine the day after for one hour and then embedded in paraffine.
The problem apperead during the cutting, because the insert started to curling up and seems that the sample is detatched from the surrounding paraffine.
How to solve this problem? What can you suggest me to change in the protocol?
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