I have lymphoid organs from mice perfunded with paraformaldehyde. The organs were cryopreserved and frozen. Could I somehow "smoothen" the fixative bonds and macerate the organ with potter (or so) to get their lymphocytes for flow citometry? I´m afraid to ruin the cells while trying to segreggate them.
You need intact cells for flow cytometry. For my opinion immunohistochemistry is the best choice for fixed and freezed tissues not flow cytometry.
You need intact cells for flow cytometry. For my opinion immunohistochemistry is the best choice for fixed and freezed tissues not flow cytometry.
Can we do flow cytometry for quantitative analysis of Heat Shock Proteins?
I wanted to see the profile of lymphocytes and I will not have fresh samples anymore... Unfortunately imunohistochemistry will not answer this question the way flow citometer would.
I do agree with @Reza You can't run flow cytometry with para-formaldehyde treated cells, we have already tried but no luck so far. As it is based for only intact cells
enzymatic separation is a common method for separate tissue cells. But this is about fresh tissue too. IHC is your only and best available method. If you could not get ideal answer from IHC, you can change or modify the IHC steps. Like change the Ab concentration or incubation time. As you know cryopreserved tissue do not need any antigen retrival.
good luck
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