I was fixing cells in coverslips with paraformaldehyde, after that I turn down the coverslip over a glass slide together with a drop of medium. We sent the samples abroad for being checked in a microscope, but they said that the cells are not viable anymore because they are dry maybe because of the transport conditions. Does anyone know how long can be cells viable? Is it better to prepare samples with immersion oil? Thank you
Hi David, PFA kills cells pretty quickly since it is membrane permeable. Oil will not help.
I think that the best way to do this is to grow cells on glass coverslips in 6 or 12-well plates and send them that way, with media.
If you are worried about shipping media, you can get a gel-like material that preserves cells during shipping from this company
http://www.ascentgene.com/products/ctm/technology/
Thank you Steingrimur. But there is a stablished protocol of action to prepare samples and fix them, indeed, even the company sent it to us. Normaly I was developing the fix with PFA in tissues. I know the killing of cells with high concentration of PFA, but it should be ok at 4%. The problem was the dead cells because of drying
Hi David, I don't know which type of cells you are working.... I work with spermatozoa and I fixed with PFA at 4% in falcon tube. If is possible, you could try to do the same and send the falcon tube to abroad. They could centrifuge it and after that throw away the supernatant and put the fixed cells on the slide to see them on microscope.
Hi David, 4% PFA is a pretty high concentration. Do you have any papers saying that cells can remain viable in 4% PFA for any length of time?
Hi, Yes, I have several papers about the treatment with that concentration of PFA in cells. I think the problema comes after that, when shipping the cells.
Thank you Laura. Our cells keep attached to the bottom of the wellplate, I mean over a side of the coverslip. So that we cannot send the cells in a falcon tube
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