[Histonet] IF-fading retardants
kmerriam2003 <@t> yahoo.com
Thu Aug 30 08:08:25 CDT 2007
Are you saying that IF stained slides can go through the normal dehydration/clearing process and be mounted in a xylene-based mounting medium?
Kim Merriam, MA, HT(ASCP)
----- Original Message ----
From: John Kiernan <jkiernan <@t> uwo.ca>
To: AGrobe2555 <@t> aol.com
Cc: histonet <@t> lists.utsouthwestern.edu
Sent: Wednesday, August 29, 2007 1:31:52 AM
Subject: Re: Re:[Histonet] IF-fading retardants
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.
----- Original Message -----
From: AGrobe2555 <@t> aol.com
Date: Monday, August 27, 2007 14:31
Subject: Re:[Histonet] IF-fading retardants
To: histonet <@t> lists.utsouthwestern.edu
> 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 C. Grobe, PhD
> International Heart Institute of Montana Foundation
> Tissue Engineering Lab, Saint Patrick Hospital
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