Cytometric Analysis of Mouse Blood

Ifor Williams, M.D., Ph.D. (irwilliams@bics.bwh.harvard.edu)
Thu, 30 Jan 1997 10:07:58 +0000

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.