Re: megakaryocytes

steven micko (steven_micko@email.eushc.ORG)
Fri, 19 Jul 96 11:24:57 EST

Julie,

We sorted rat bone marrow megs in 1980-81 at Washington Univ in St.
Louis. We ran the marrow through the Beckman Elutriator to concentrate
the megs about 150-fold, then stained with Hoechst 33342 and sorted on
a FACS IV for the different ploidy cells. In one application, we
sorted directly into a drop of glutaraldehyde on microscope slides and
then stained with acetylcholinesterase and Wright's stain. We observed
that the higher the ploidy we tried to sort, the more fragile were the
megs -- apparently they were beginning to shed platelets. We used an
80 micron nozzle with very few plugging problems. Light scatter was
unremarkable.

Is this the kind of information you need? Let me know if you want to
discuss this further, viz. staining conditions, metachromatic effects
of HO33342, etc.

Best regards,

Steve Micko
Emory Hospital
Atlanta

P.S.
I stood outside the hospital for two hours yesterday to see the torch
bearer run by in about 15 seconds. Let the games begin!!

______________________________ Reply Separator _________________________________
Subject: megakaryocytes
Author: Julie Auger <jauger@flowcity.bsd.uchicago.edu> at INTERNET
Date: 7/18/96 7:22 PM

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From: Julie Auger <jauger@flowcity.bsd.uchicago.edu>
Subject: megakaryocytes

Hi folks- Any experience with flow and megakaryocytes? Any info would be
appreciated with regard to light scatter characteristics or fluorescent probes.

Thanks

Julie
********************************************************************************
Julie A. Auger voice: 312-702-9212 lab
Director, Flow Cytometry Facility 312-702-9261 office
University of Chicago fax: 312-702-3701
5841 S. Maryland Ave. MC-1089
Chicago, IL 60637 e-mail: jauger@flowcity.bsd.uchicago.edu
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