My previous message was mostly deleted so am resending: Lora Barsky asked: "Can anyone offer insight as to why my users have been flow problems when using 293A and 293T cell lines on BD FACScan and BD FACSCalibur? The cell lines are adherent, so they trypsonize to release from the plates, they bring the cells in DPBS to FACS. They're all looking for GFP expression." ------------------------------------------------- DPBS contains calcium and magnesium, which usually accelerate clumping. You might try adding EDTA or use Ca and Mg free PBS. Also keeping the cells cold will reduce clumping. Finally, many clumps are due to dying cells releasing DNA that traps other live cells. Adding a small amount of DNAase can solve this problem. Good luck, Tony Bakke Antony Bakke, PhD Director, Immunology & Flow Cytometry Oregon Health Sciences University bakkea@ohsu.edu
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:53:55 EST