Re: Xylene substitutes

From:Gayle Callis <>

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 AND
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
>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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