From: Steve G. Hilliard (steve@habanero.cb.uga.edu)
Date: Thu Jan 28 1999 - 10:43:38 EST
Keith,
I always warn my clients about exposure to light, but I did directly test
some FITC CD4/8 staining and saw no loss of signal. We ran the samples,
put them in a brightly lit window for about 2 hrs, and ran them again, and
there was no discernable effect. Funny, but I still caution them about
exposure with PI.
Just one data point....
-------------------------------------------------------------
Steve G. Hilliard flowman@uga.edu
University of Georgia Cell Analysis Facility
http://floweb.cb.uga.edu/Floweb/
-------------------------------------------------------------
On Tue, 26 Jan 1999, Keith Bahjat wrote:
>
> This message brings up a point that I'd like to see addressed by the experts.
> In my hands, FITC (as well as PE, PerCP, APC, PI, 7-AAD, and ToPro-3) is very
> stable, and no precautions need to be taken while staining or running samples,
> as no degradation has ever been seen. Every clinical and research lab I've been
> in stains samples on the bench top and takes no special precautions to avoid
> any forms of light. But some researchers here do things like staining in the
> dark, encasing samples in aluminum foil, and using red filters on the room
> lights in the flow lab for fear of fluorochrome degradation. Based on my
> experience, I'm not sure why they fear the light to this extent. I've seen this
> happen using the fluorescent scope, but I think those are usually fluors like
> Cy2 and Cy3. Are some derivations of fluorescein more photolabile than others,
> or am I ruining my samples.
>
> thanks for any news and views.
>
> Keith Bahjat
> kbahjat@ufl.edu
>
>
> joan Kalnitsky wrote:
>
> > Does anyone know the quenching time of PI? We have someone who wants to
> > stain with PI and is concerned about whether it will fade out like FITC
> > would. Also, how about PE. Is it as light sensitive as FITC. Any and all
> > suggstions would be helpful. If anyone has stained with PI and has a
> > protocol, we would love to see it.
> > Thanks in advance.
> > Joan K
> >
> >
> >
> > Flow Cytometry Lab Supervisor
> > VMRCVM
> > (540) 231-4115
> > FAX 540-231-7367
> > jkalnits@vt.edu
> >
> > "It is better to serve than to receive."
> > B. Borg
>
>
>
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