Flowcytometri on PBMCs (frozen) CD4+, CD8+, CD25+, CCR7- is this possible? - (Jun/18/2012 )
I'm planning a small FC study. I want to use peripheral blood and use a density gradient to seperate into MCs (Ficoll Paque Plus). My plan was to freeze down these PBMCs at -80 Celcius.
I'm going to gate the different subtype (CD4+/CCR7- and CD8+/CCR7-), but then I started to wonder if this was possible to freeze the cells before running them on a Flowcytometer. Do you have any experience in this when using the above antibodies?
I have stained frozen PBMC for CD4, CD8 and numerous other markers and it works just fine. About the only problem you may have is clumping of the cells after you thaw them and the clumps can clog the cytometer. If you add DNase (10 U/mL) it will prevent clumps.