In message  <[log in to unmask]> Confocal Microscopy List writes:
> In a message dated 12/5/97 15:18:59, [log in to unmask] wrote:
>
> <<Has anyone had experience with antifade reagents, in particular
> Molecular Probes ProLong, SlowFade, and SlowFade Light. I know it makes
> no logical sense, but in our experience fluorescent labels of rat tissue
> fade under the laser a whole lot faster than in human tissue.>>
>

I dont know if this helps or whether it has anything to do with the human vs rat
observation, but I was advised by someone at Leica that the freshness of the
prep was really important. The fresher the prep the bigger the problem with
bleaching. Maybe your rat tissues are taken and used sooner post mortem than the
human stuff? Anyway,one recommendation which was made to me to reduce bleaching
was to leave the mounted specimens at least a day before viewing. I guess as
with most things there will be a trade off against the deterioration of the
specimen with time. I use mowiol so I have to leave the preps at least overnight
to set. From what I understand, bleaching is all a problem of oxygen and
oxidative free radicals, therefore one of the things which I do is to degass the
mountant! I have never directly compared degassed vs normal, but I am in general
very happy with the stability of fluorescence in my coverslips. With mowiol
there is of course the problem of the cells becoming a bit flattened by the
compression of the mowiol when it dries, but as I do not do 3D reconstructions,
this minor irritation is more than compensated for by the ease of use and
stability.
I hope this is some help
Matthew

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     Dr. Matthew J Hannah
     Institute for Neurobiology
     University of Heidelberg
     Im Neuenheimer Feld 364
     D-69120 Heidelberg
     Germany

     tel: +49 (6221) 54-8320 or 54-8692
     fax: +49 (6221) 54-8301

 http://server.nbio.uni-heidelberg.de/neurobiology.html
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