Dr. O'Malley,

I have used Annexin V-FITC (from Pharmingen) to do live imaging of
apoptotic cells in brain slices.  Annexin V binds to phosphatidyl serine
residues which get "flipped" to the outside of the membrane during
apoptosis.  Unfortunately, the Annexin V cannot penetrate deep into the
tissue so this technique is mainly useful for imaging cells on the
surface.  You might be able to modify the incubation procedure to get a
little more penetration.
Regards,

Tarik


Donald O'Malley wrote:

> Hi All,
>
>     We are trying to stain dead/dying cells in living zebrafish.
> We would like to do this non-invasively, i.e. by simply incubating
> the larval fish in the "dead-cell" stain.  We've tried propidium iodide
> over a range of concentrations and for times from several hours
> to overnight, all to no avail.  Presumably it can't diffuse into the
> fish, although we know that some compounds are able to diffuse
> into the fish.   I realize this is a bit esoteric, but any suggestions
> would be appreciated.   I also know there are live-dead kits from
> Molecular Probes, but before we spend hundreds of dollars, I
> thought I might check first with the imaging community.  (We are
> gently killing the cells with a low-intensity laser beam; we want
> to confirm cell death before doing extensive behavioral testing).
>
> Thanks,
> Don
>
> Donald M. O'Malley, Ph.D.
> Department of Biology
> 414 Mugar Hall
> Northeastern University
> Boston, MA 02115
> [log in to unmask]



--
Tarik Haydar, Ph.D.
Yale University Medical School
Section of Neurobiology
Sterling Hall of Medicine, C-319
333 Cedar Street
New Haven, CT 06520

Phone: (203) 785-5418
Fax: (203) 785-5263
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