Re: Re:[Histonet] IF-fading retardants

From:John Kiernan

Why do you "suspect you would kill your signal" by dehydrating, clearing and  mounting a slide carrying a fluorescently labelled antibody? 
Alcohol coagulates proteins on-the-spot, complete with any covalently bound fluorescent tags. This was established for lectin histochemistry 30 years ago, and has been well documented for fluorescently labelled antibodies in more recent years. The intensity of  fluorescence emission may be less in DPX than in buffered glycerol, but who has done comparisons? 
A fairly recent study strongly favours alcohol dehydration, clearing and mounting in a non-flourescent  recinous medium. for permanent immunofluorescence slides.
John Kiernan
Anatomy, UWO
London, Canada
----- Original Message -----
Date: Monday, August 27, 2007 14:31
Subject: Re:[Histonet] IF-fading retardants

> Carl,
> I used to use "FluorSave" mounting medium from Calbiochem (Cat 
> #  345789).  
> This mounting medium "hardens" so the coverslip doesn't 
> move  and you don't 
> have to seal around the edges with nail polish.  It also 
> kept  the signal from 
> fading.  I still stored the slides horizontally at 4C 
> or  -20C.  I wouldn't 
> suggest try in to dehydrate through xylene and  
> coverslipping, as I suspect you 
> would kill your signal.  
> Albert
> Albert C.  Grobe, PhD
> International Heart Institute of Montana Foundation
> Tissue  Engineering Lab, Saint Patrick  Hospital
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