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Hi,

I am using the succinimide ester of bodipy dyes (Molecular Probes) to
label my neutrophils and it appears to do a good job of nonspecifically
labeling the whole cell (live).  I dilute it, toss it on, wait 5
minutes, then wash and centrifuge twice.  The neuts appear to remain
functional, gobbling up stuff and spreading and I am positive that I am
seeing the whole cells.   I can then fix and stain with a phalloin
conjugate and get a pretty picture of actin filaments as well.  I expect
that if you wanted to you could try it on already spread cells and
gently rinse the excess off.

If you want I could come to Spain and help you.  I love your wines.

Deb Berglund
Montana State University

F Javier Diez-Guerra wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear confocalists,
>
> I am studying cell spreading on fibronectin-coated coverslips and would
> like to know about fluorescent probes that could be used to measure
> cell area.
>
> Ideally, the probe should faithfully delineate the cellular contour, give
> enough signal/noise ratio for easy thresholding and, if possible, be used
> after (PFA) fixation.
>
> We also would like to be able to combine it with conventional
> immunofluorescence.
>
> Thanks.
>
>
> F Javier Diez-Guerra, PhD
> Profesor Titular
> Centro de Biologia Molecular Severo Ochoa
> Facultad de Ciencias
> Universidad Autonoma
> Cantoblanco, 28049 Madrid
> SPAIN
>
> phone:  +34 91 4978051
> Fax:      +34 91 4978087
> e-mail: [log in to unmask]
>
>
>