Macrophage detached - (Apr/03/2001 )

Dear SirI am currently using macrophages (5-7 days old) to run flowcytometery on these cells. The first step to start with is to detach cells from plate/flask. Would you please tell me what is the best way to detach these cells without any disturbance to the cell surfceusually I am using cold 0.02% of EDTA and 0.25% trypsine and still I have difficulties to lift these cells up in short time beside lose almost 30% of cells.Many thanks for your help.

M. AlaliCentre for Virus ResearchNSW, Australia

To detach macrophages you could use lidocaine (4 mg/ml) togetherwith 5 mM EDTA. leave them at 37C for 3-5 minutes and they should come off.