We are currently facing the problem that we can't do FACS analysis on isolated cells because they clot together after doing lavage with PBS and 3%FCS. How can we avoid that?
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Alexandre Borbely
Try adding 1 or 2 mM EDTA in your PBS. It should prevent cell clots.
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Jason Gardner
EDTA might help. We use 0.6mM EDTA to increase the # of cells isolated from lung lavage.
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Evan Chiswick
Yes, I'm well acquainted with the pains of trying to recover cells post-CLP, and then using the cells for functional assays! We do not use FCS for lavage; cold Hbss or PBS with 1mM EDTA or 10U/mL of heparin works fine. Strain the cells through a cell strainer to help remove cecal contents. Wash with Hbss/PBs with 0.5% BSA. There are ways to get rid of the bacteria and dead cells too.
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