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Dye taken up prior to fixation will most likely be rendered unusable by 
(1) leakage through protein "holes" (in membranes) created by the
fixation 
or 
(2) reaction with the anilinium nitrogen.  
But you might want have to try.  One indicator of membrane holes is the
lack-of-punctatedness of staining of aldehyde-fixed and then washed
tissue infiltrated with dye.
The dye bleaches noticeably at 100 W/cm2 illumination.  But what keeps
the dye from bleaching at the fast rate of fluorescein et al is the low
extinction.

Walter Niles

-----Original Message-----
From: Amitabha Majumdar [mailto:[log in to unmask]] 
Sent: Wednesday, May 01, 2002 10:48 AM
To: [log in to unmask]
Subject: FM1-43 and fixation.


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I would like to know whether one can image FM1-43 dye uptake after
fixation.If yes what type of fixation will help.I am trying to do the
experiment on drosophila neuromascular junction by stimulating using
high K.Does it bleaches very fast.


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                  Amitabha Majumdar.
                   Research Scholar
            Department of Biological Sciences.
          Tata Institute of Fundamental Research.
                 Homi Bhaba Road,Colaba
                    Mumbai-400005
                       India.
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