I am doing flow analysis on blood from transgenic mice carrying oncogenes that are expected to predispose to the development of myeloid lineage leukemias. I have two questions for those who have experience with analyzing mouse blood specimens: 1. What erythrocyte lysing solutions work well with mouse blood? I have used a lysis buffer containing 0.16 M NH4Cl, 0.1 mM EDTA, and 12 mM NaHCO3 (5 to 10 minute incubation at room temperature followed by return of the cells to standard staining buffer). It works reasonably well, but occasionally there are more residual erythrocytes in the samples than I would like to see. I am interested in the experience of others with commercial lysing solutions or other home brew lysing reagents for use on mouse blood. 2. Is there a convenient method to analyze platelets (number and FSC/SSC characteristics) in heparinized whole blood from mice? Any suggestions on instrument settings (we are using a BD FACScan) or references would be appreciated? One manifestation of some of the leukemias in these mice may be abnormalities in development of the megakaryocytic lineage. Thanks in advance for those who can share some tips in this area.
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