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Dear Dale--

I know of no way to selectively quench the fluorescence of Alexa 
fluorochromes.  However, back in the 1980s, we'd occasionally use 
elution-restaining to multi-stain tissue if we needed to localize two 
different antigens with primary antibodies raised in the same species.  
It involves treating the sample with potassium permanganate and 
sulphuric acid--not gentle, and background goes up.  If you're stuck, it 
might provide an approach.  A number of authors have used it to elute 
fluorescently labeled antibodies.

Here's the reference:
Tramu,  G.,  A.  Pillez,  and  J.  Leonardelli  (1978)  An efficient  
method of  antibody  elution  for  the  successive or  simultaneous  
location  of two
antigens  by  immunocytochemistry.  J.  Histochem.  Cytochem. 26:  322-324.

Good luck!

Martin




On 2/16/2017 10:12 AM, Dale Moulding wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images onhttp://www.imgur.com  and include the link in your posting.
> *****
>
> Dear all,
> does anyone know of a way to irreversibly destroy alexa fluor fluorescence so a sample can be stained with a second set of dyes?
> We do whole mount staining for 3 antigens with Alexa 488, 568 and 633. We would then like to re-image the same samples with cellmask and phalloidin.
> Is there any way to kill alexa fluor fluorescence without destroying membranes and phalloidin binding?
> Cheers
> Dale

-- 
Martin Wessendorf, Ph.D.                   office: (612) 626-0145
Assoc Prof, Dept Neuroscience                 lab: (612) 624-2991
University of Minnesota             Preferred FAX: (612) 624-8118
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