Re: RBC analysis

From: Robin Barclay (Robin_Barclay@compuserve.com)
Date: Fri Feb 18 2000 - 05:30:09 EST


Message text written by INTERNET:David.McFarland@mcmail.vanderbilt.edu
>Hello everyone.  I have a client that wants to do analysis of RBCs.  This
is new
to me.  Seems everyone else wants to make sure we avoid the RBCs! 
Basically,
they are injecting mice with a FITC-labeled ligand that should only be
bound
(and internalized?) by cells that express the receptor for said ligand. 
They
have control mice, and transgenics that express the receptor.  The question
is
where is it expressed?  We did some earlier experiments excluding the RBCs,
but
now they want to look at them as well.  They are looking to me for advice
and I
don't have any to give them.  So, does anyone have some good advice to pass
along?  I'll even give you the credit!
David McFarland<

We often study human RBC (and platelets) as well as leucocytes.
RBC are predominantly slightly smaller than lymphocytes and of low
granularity (for locating in FSC/SSC profiles) but this is heterogeneous
and varies.
RBC are 100 - 1000 fold more numerous than leucocytes, so will predominate
in any whole blood sample - so scale your sample size accordingly.
Obviously - don't lyse.
Since there will be size/granularity overlap with leucocytes use another
marker to discriminate - such as glycophorin A for RBC or CD45 for
leucocytes.
I don't know what is equivalent for these in mice - but I hope this gives
some ideas.
Robin Barclay
SNBTS Edinburgh



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