Anita, The unfortunate thing about using mouse whole blood is that it frequently does not lyse completely. This does not normally interfere with the staining, it only makes acquisition a little more challenging. What size tubes are you lysing in? What volume:volume ratio are you using? Randy T. Fischer NIH/NIAMS Building 9 Bethesda, MD 20892 fischer1@mail.nih.gov (301) 594-3537 > ---------- > From: Haugan, Anita > Sent: Thursday, November 16, 2000 4:35 AM > To: Cytometry Mailing List > Subject: NH4Cl lysing of murine whole blood > > > To all of you who use ammonium chloride lysing of RBC in whole blood > samples > from mice for surface markers: > > I need some advice concerning lysing of murine RBC. I have just started to > work with whole blood from BALB/c mice, and are trying to lyse the RBC > with > a NH4Cl-solution. My problem is that my samples aren't completely lysed, > and > one can see that the pellets are redish in some cases. My qustions are: > > What is the optimal g-value for centrifugation of the samples to get > maximum > lysis? We use 1200rpm (270g) for 10 min. > > Is there anything else we should be aware of in using NH4Cl lysing - both > concerning the lysing method and analyzing the samples on the > flowcytometer? > > So if anyone have some answers/advices or best of all a detailed protocol > of > how to lyse RBC in murine whole blood they could send me, I will be > extremly > happy! > > Thanks in advance! > > Anita Haugan > Department of Vaccinology > National Institute of Public Health, Norway > >
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