Re: Murine embryonic heart cells

From: Mark Kukuruga (kukuru@med.umich.edu)
Date: Thu Nov 08 2001 - 14:33:48 EST


Janet,
Assuming this is a total organ digest . . . you probable see a "smash" of cells,
rather than populations.  It's entirely possible you're not looking at cells at all,
but rather mostly bits and clumps (with some cells thrown occasionally as a teaser).
Have you view the samples through a microscope?  This can be very revealing, and may
give you the answers you seek.
Also, think of a DNA stain (PI, 7-AAD, Hoechst, DAPI, others) . . . you can then
trigger on the stained cells, and use that to limit your analysis to intact cells.
It may help you home in on the pop. of interest.
MAK.

--
Mark A. KuKuruga, Managing Director
University of Michigan Flow Core
7416 CCGC 0946
(734) 647-3216, fax (734) 936-7376
kukuru@umich.edu


>>> janet dow <jldow@unity.ncsu.edu> 11/06/01 04:12PM >>>

I hope that someone can help me with this problem as I am somewhat at a
loss as to what to tell my client.
He wants to do flow on embryonic mouse heart cells.  He is staining for
myosin heavy chain and for apoptosis with tunel.  The problem is that I am
unable to identify distinct populations(using forward and side scatter)
within the cell suspension he is providing to me, and I am concerned that I
am missing the important cells.  I have tried looking at the ungated data
for the myosin staining and have come no closer to finding populations of
cells.  I know that it is a mixed population of cells but expected to be
able find populations with the mix.  Another problems is that of course the
hearts are really tiny and he is pooling many hearts together.

I really appreciate any help anyone can give me with this situation.

thanks in advance.

Janet Dow

Janet Dow
Research Technician and Manager
Flow Cytometry Facility
North Carolina State College of Veterinary Medicine
Room C-314
Raleigh, NC 27606
(919)513-6364



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