From: Mike Daws (micdaw@hotmail.com)
Date: Tue Sep 22 1998 - 13:05:36 EST
Dear Carine You may like to try Pucks Saline with EDTA (0.5mM I think) which I have used in the past for removal of adherent cells, and which I found to be far less traumatic to the cells than a PBS/EDTA removal. I have no idea why Pucks saline should be so much better than PBS for this, but for me the difference was rather dramatic (>95% survival vs approximately 60% survival with PBS/EDTA) and reproducible. This may not be the answer in your case, but it may be worth trying. Good luck, Mike Daws >Dear Flow-ers, > >We started to study apoptosis 4 years ago. >We're working with an adherent cell line and we're using flow cytometry >detection of propidium iodide stained cells after ethanol fixation >(apoptotic cells = sub-G1 peak). But we have lots of problems because of >this cell line (which is adherent !), and the cell detachment is a very >delicate step : we need to save living cells as well as apoptotic cells >(and apoptotic bodies). > Now we're using PBS/EDTA buffer with a very low addition of trypsin, but >it's not the absolute solution. So we're looking for ideas in order to >simplify this step or to substitute the method. >Thank you for any help ! > >Best regards, > >Carine MALCUS > > >__________________________________________________ > > Carine MALCUS-VOCANSON, Ph. D. > // cellular biology // bioethics // > > bioMerieux S.A. > UMR 103 CNRS - bioMerieux > E.N.S.L. > 46, allee d'Italie > 69364 LYON cedex 07 > FRANCE > > Ph : 33 (4) 72 72 85 92 > Fax : 33 (4) 72 72 85 33 > email : Carine.Malcus-Vocanson@ens-bma.cnrs.fr >__________________________________________________ Maryalice Stetler-Stevenson Director Flow Cytometry Unit Laboratory of Pathology, NCI, NIH <<<<<<<<<<<<<>>>>>>>>>>>>> Michael R. Daws, PhD Immunology (111R) VA Medical Center 4150 Clement St San Francisco CA 94121 Tel: 415-750-2104 Fax: 415-750-6920 E-mail: mdaws@itsa.ucsf.edu <<<<<<<<<<<<<>>>>>>>>>>>>> The most thoroughly wasted of all days is that on which one has not laughed........Chamfort ______________________________________________________ Get Your Private, Free Email at http://www.hotmail.com
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