RE: Xylene substitutes

From:"Tarpley, John" <>

It is fairly easy to test the compatibility of your mounting media with a
xylene substitute of the nonlimonene variety. Simply place a few drops of
the substitute on a slide, add mounting media, and a coverslip. If
everything mixes well and dries clear the two are compatible. If a
precipitate begins to form then they are incompatible. We use Shandon's
mountant because we find it has the least objectionable odor of the ones we

John Tarpley  5-1-A
Associate Scientist,
Amgen Inc
One Amgen Center Drive
Newbury Park, CA 91320

> ----------
> From: 	Gayle Callis[]
> Sent: 	Tuesday, September 26, 2000 4:21 AM
> To:
> Subject: 	Re: Xylene substitutes
> Good advice from Sarah!  Try the mounting media sold by Anatech that is
> compatible with Propar, Shandon and Richard Allan have compatible mounting
> media.   We use the Richard Allan and it is compatible with both xylene
> Clearite 3 or Propar, never had a bleeding problem with any stain combined
> with either these xylene substitutes or xylene.   
> >
> >Dear Mary,
> >   I'm not familiar with Refrax mounting media, but if it is a clear
> media, it is not compatable with Pro-Par.  You have to use one of the
> yellow medias like Coverbond or Permount.  The mounting media is making
> the
> stains bleed.  I have been doing Trichr
> >
> >Sarah Christo, HT (ASCP)
> >Research Associate, Histology Lab
> >Texas A&M University
> >College of Veterinary Medicine
> >Dept. of Vet Anatomy & Public Health
> >College Station, TX  77843-4458
> >phone: (979) 845-3177
> >fax:  (979) 458-3499
> >
> >>>> "Hess, Mary" <> 09/25/00 02:24PM >>>
> >I've received some responses asking why we used xylene for Masson's
> >trichrome instead of pro-par in the first place. Initially it was because
> I
> >got the Masson's Trichrome protocol and the kit from another person at my
> >company who was very much into using only 'pure' reagents (ie, expensive
> >absolute alcohols, xylene, etc) to ensure the quality of his slides. And
> the
> >quality is very good when you use those solutions. When I started, I
> quickly
> >switched to our normal alcohol and found no difference between his slides
> >and mine. I have now tried to do our Masson's trichrome using pro-par and
> >refrax mount instead fo xylene and permount. My initial results aren't
> too
> >good. 
> >Recently we have found eosin 'bleeding' from our H&E tissues when the
> slides
> >were mounted (for which we used Pro-Par and Refrax mounting media). I
> >switched out everything (from water, to alcohol, to Pro-Par, to eosin and
> >hematoxylin) in our H&E stations and still had the problem. When I did
> the
> >trichrome with the Pro-par,  the Biebrich Scarlet Acid Fuscin bled into
> the
> >mounting media. I don't know what it is. If you guys do I would be
> grateful.
> >And in the end, it was the guy who started me on the stain who kept me
> using
> >xylene. I don't want to keep using it, but so far, it seems like I'm
> going
> >to have to. I hope this 'bleeding' is something I'm doing wrong and is
> >easily fixable, I'd rather it be my fault. Thanks for the help so far....
> >-Mary Hess, San Diego, Ca.
> >
> >
> >
> >
> >
> Gayle Callis
> Veterinary Molecular Biology
> Montana State University
> Bozeman MT 59717-3610
> 406 994-4705
> 406 994-4303

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