[Histonet] fluorescence with xylene dehydration

[John Kiernan]

Yes, I've done this with fluorescein- and rhodamine-labelled protein tracers and lectins. The fluorescence is still there years later, in slides stored in boxes at room temperature. 
 
Permanent mounting media are also OK for preparations stained with fluorescent antibodies. See Espada J, Juarranz A, Galaz S, Canete M, Villanueva A, Pecheco M & Stockert JC (2005) "Non-aqueous permanent mounting for immunofluorescence microscopy" Histochem. Cell Biol. 123: 329-334. A Scopus search shows 7 papers citing this one (2006-2009), all in good journals
 
Alcohol dehydration of the sections coagulates the fluorescently labelled protein in situ. After clearing, it's important to use a non-fluorescent resinous mounting medium. In my experience DPX (polystyrene based) is OK and so are at least two of the poly(methylmethacrylate) media, Entellan and Cytoseal. Canada balsam is unsuitable because it fluoresces enough to interfere with interpretation of the slides.
 
John Kiernan
Anatomy, UWO
London, Canada
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----- Original Message -----
From: Emily Sours <talulahgosh <@t> gmail.com>
Date: Monday, February 1, 2010 13:19
Subject: [Histonet] fluorescence with xylene dehydration
To: histonet <@t> lists.utsouthwestern.edu

> Hello
> 
> Has anyone used a xylene dehydration at the end of fluorescent 
> staining?I've searched the histonet archives for this answer but 
> the answers were
> mixed.  I thought I'd ask again to see if there were any 
> new opinions.
> 
> Emily
> 
> 
> 
> ...the thrill of being close to that hidden knowledge. That's 
> the way I feel
> when I read Nabokov. Encrypted within his words, encoded 
> indecipherably,ambiguously, is the equivalent of the secret of 
> lightning. Something akin to
> the secret code of higher human consciousness, the DNA, the 
> genome of
> genius.
> -Ron Rosenbaum
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