I need to fix whole blood and splenocytes from mice and save for flow cytometry staining for another day. One of my antibodies is sensitive to parafilm fixation and needs to fix using acetone. How long will cells be okay fixed with acetone, resuspended in PBS after, and left in the four-degree fridge?
Dear Misha Fini
If you are going to perform the flowcytometry then i suggest not to go with fixation. keep the sample in 8-15 degree and perform staining next day.
Although, Acetone fixation is not practically perfect in flowcytometry as this cause the shrinking of cells. People prefer fixation with chilled methanol or Para-formaldehyde (PFA). Acetone fixation is good and being performed in routine practice of ICC or IHC.
I hope this is helpful.
Good luck.
Jitendra
Article Flow acetone-staining technique: A highly efficient procedur...
Hope this will help.
Hello Misha, If you have fixed the cells in PFA overnight, then they can be spun and resuspended in PBS for staining. Of course it depends on whether the antigen is on the surface or intracellular as to what method you would use.
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