Brian - I have had pretty good luck in the past on both human & mouse whole blood with BD's FACS Lyse using similar whole blood staining techniques. It does contain a fixative, though - Clare Rogers Applications Scientist Accuri Cytometers, Inc. 173 Parkland Plaza Ann Arbor, MI 48103 734-994-8000 x120 734-994-8002 fax crogers@accuricytometers.com www.AccuriCytometers.com ________________________________ From: Brian McFarlin [mailto:bmcfarlin@mail.coe.uh.edu] Sent: Sun 2/10/2008 11:47 PM To: cyto-inbox Subject: Need Help with Preparing Mouse Blood Leukocytes I have a unique situation and I need some guidance. We are trying to label 5 uL of whole blood (Heparin treated) collected from the saphenous vein (we actually collect 30-40 uL and then split it up). We ran though this last week and used RBC lysis buffer from a company called eBioscience. We used 400 uL of RBC lysis buffer with 10 uL of stained cells (5 uL blood, and antibodies to 10 uL). We lysed for 5 minutes and found that when we tried to analyze the samples, we just got what looked like a bunch of RBCs on the FS vs. SS plot. We concluded that it is either actually RBC contamination or we killed all the cells during the lysis step causing us to see only debris. We have previously used this exact same procedure with humans without any problems. We are able to make measurements on these small blood samples because we are using a Gauva EasyCyte Mini. Any suggestions that anyone has would be extremely helpful as we do not have extensive experience with mouse blood samples.Received on Tue Feb 12 14:18:00 2008
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