[Histonet] Goldner's Trichrome

Gayle Callis gcallis <@t> montana.edu
Tue Sep 18 14:30:52 CDT 2007

Dear All,

The monochromatic effect could also be a problem since it is being done on 
thick, ground bone sections embedded in Methyl methacrylate.  It would be 
better to microtome with tungsten carbide knives to obtain thin, 5 um 
sections,then remove the MMA completely with hot xylene or a suitable 
solvent, then do the Goldners.

Methyl methacrylate is extremely hydrophobic and large molecular weight 
dyes cannot penetrate or if they do, it creates havoc with the 
staining.  Richard Horobin wrote a wonderful article about the effects of 
staining on plastic sections some years back.   (Sorry, reference is at 
home). He even discussed how Masson trichrome staining can be reversed 
(something some of our bone experts noticed on the on when doing this stain 
on plastic sections) so that blue collagen is red and the collagen stains 
red instead of blue with aniline blue.

  Surface staining is usually done with dyes like methylene blue/basic 
fuchsin, toluidine blue at pH 8, MacNeals tetrachrome, and Sandersons rapid 
bone stain from Surgipath which is a potassium permanganate oxidized 
methylene blue mixture, a variation of Stevenels blue.  Ms. Sanderson 
(Mayton) figured out an easier way to make the staining solution.

Mild acid etching makes this kind of staining even better since it removes 
a few um of calcium from surface of exposed bone so one can have better 
depth of dye penetration.    All dyes used in these methods are of low 
molecular weight and can penetrate the plastic better with heat, high pH, 
and after etching.

We preferred to do MacNeals tetrachrome for connective tissue staining on 
ground MMA bone sections, but is also is brilliant with toluidine blue 
added. Basic fuchsin can be used as a counterstain with toluidine blue and 
Sandersons bone stain.  You very quickly learn what is connective tissue 
versus bone tissue components, since much of the staining is in shades of 
blue or green depending on what stain is used.  Unfortunately, MMA embedded 
bone will present some problems for the ground thicker sections as staining 
is a bit limited as compared to microtomed MMA sections with plastic 
removed or decalcified bone embedded in paraffin.  It is challenging but 
fun too.

  At 09:37 AM 9/18/2007, you wrote:
>See Fred Monson's Histonet reply (undated):
>It looks like a slight variant of Masson's trichrome; none of the dyes 
>used in it should give metachromatic effects.
>John Kiernan
>Anatomy, UWO
>London, Canada
>----- Original Message -----
>From: "Malinowski, Joanne O," <Joanne.Malinowski <@t> crl.com>
>Date: Monday, September 17, 2007 8:46
>Subject: [Histonet] Goldner's Trichrome
>To: histonet <@t> lists.utsouthwestern.edu
> > Hello Plastic Histonetters,
> >
> > Can anyone share the latest Goldner's Trichrome for MMA ground
> > sections?Ours turns out monochromatic. If not Goldner's, what is
> > your choice for
> > connective tissue for ground MMA sections?
> >
> > Thank you.
> >
> > Joanne
> >
> > Joanne Malinowski,HT ASCP
> >
> > Plastics Lab Manager, Medical Devices Division
> >
> > Charles River Laboratories
> >
> > Pathology Associates
> >
> > 15 Worman's Mill Court, Suite I
> >
> > Frederick, Maryland 21701
> >
> >
> >
> > Phone 301-624-2034
> >
> > Fax 301-663-8994
> >
> > Email: joanne.malinowski <@t> us.crl.com
> >
> >
> >
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610

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