Any one working with mouse blood?

From: Barren, Phil <BarrenP@MedImmune.com>
Date: Tue Sep 07 2004 - 14:08:58 EST
FLOWers

	I feel humbled to ask, but is anyone successfully immunophenotyping mouse blood? 
What I would like to do is draw the blood, block the Fc receptors and then stain (direct)
for Lymphocyte markers and ultimately lyse.  In the end I would like 100% viable cells. I
want to be able to use the blood scatter populations to help me determine the gates for 
immune cells in lung

I have stained and lysed with FACSLyse, and the scatter patterns look as expected, but
the cells are fixed.

I have tried other commercial (viable) lysing reagents, but end up  with odd scatter,
degranulated granulocytes (in my lymphocyte gate CD45 confirmed) and low viability.

The manufactures have suggested  heparin or EDTA, and I have tried both.

Of note these viable lysing reagents work just fine with human blood, but the mouse has
not worked out. 

Any help would be greatly appreciated.

Thank you for your time

Pb
Received on Wed Sep 8 12:38:00 2004

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