Hello all,
my question is about using my frozen samples to detect macrophages. I have uterine cells, scratched with 40um strainer and frozen in -81 degrees in 90% FBS and 10% DMSO medium. They was centrifuged with 1500 rcf. My question is, can I use this samples to detect macrophages? It will be very usefull for me to mark them in this samples for flow cytometry, but I dont really know if it have any sense since this samples was frozen for other purposes and it is propably not most optimal way to freeze it for macrophages.
Thank you for all answers,
Szymon
My advise would be to check once for macrophages (frozen cells versus fresh samples) with CD64 and not only CD14 to be sure that the receptor was not internalized during the procedure
Regards!
The freezing medium for macrophages isn't any different from what was used. But the storage at -80C is the concern. Since you don't need these cells to be revived...it might be possible that the cell membranes are intact for flow staining (after the thawing process). If you can get fresh samples, that would be the comparison as stated above. But if you can't get fresh tissue samples for comparison...you should still be able to determine intact cells versus other stuff. And as stated above, use multiple markers...just in case...
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