I'm preparing the samples for my experiment for TEM. I did the resin embedding step. After adding the pure resin I centrifuged the cells at 1,500 rpm 5 minutes but could't pellet it well. The technician told it is because I didn't use fresh resin. I prepared the resin yesterday and left in the fridge, today I thawed it and used. After centrifugation at 2,000 rpm for 7 minutes, they were still suspended in the resin. Is there any way to save my samples?
try to heat the resin, it will be more fluid so probably you can spin down your cells. For the next embedding spin down the cell in low melting agar before starting the dehydration, start the whole procedure with the agar blocks (also easier to handle) and no problems with spinning down at the end.
Dear Gerda
I tried heating the resin and centrifuged at 2,000 rpm for 7 minutes but still the cells didn't settle down.
Shaima
If:
1. Your previous steps in specimen prep (dehydration, resin infiltration) were performed properly
2. Most of your suspended cells are in the bottom part of vial
Then you can proceed with polymerization. You will have enough cells on your sections to observe them.
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