Re: Sensitivity to room light.

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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