Cytometric Analysis of Mouse Blood

From: Ifor Williams, M.D., Ph.D. (irwilliams@bics.bwh.harvard.edu)
Date: Thu Jan 30 1997 - 05:07:58 EST


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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