[Histonet] Re: Hepatocytes trichrome

Gayle Callis gcallis <@t> montana.edu
Mon Jul 12 13:53:19 CDT 2004

It is important to remember Massons trichrome stains ALL connective tissue
components, collagen, reticulum fibers and basement membranes.  If you want
collagen only, you should do Van Giesons stain. 

You did not say which trichrome method you are using, the classic Massons

IN Massons trichrome, the Biebrich scarlet - acid fuchsin step can be
controlled by 

1.  	Less staining time in Biebrich scarlet/AF solution, if your method
says 5 min, try 1 min.  

2.  	Phoshptungstic acid/phosphomolybdic acid step in controlled with
microscope until stain is   	differentiated out of cells.  It may take
longer than time stated in your protocol for PTAH/PMA (	his solution shoule
be fresh each time). 

OR one can simple eliminate the biebrich scarlet/acid fushsin, just do
aniline blue alone with Weigerts iron hematoxylin. If you don't need to see
muscle or red blood cells, just take away the red! This leaves connective
tissues blue with darker staining nuclei.  

We used the Richard Allan trichrome kit last week for the first time and
had the kind of staining you describe with nice staining on lung but
without overstaining of cells other than muscle.  Red Blood cells were
spectacularly red, however. It could be the timing of reagents developed
for their kit that makes it work nicely.  We did let sections sit in Bouins
overnight at RT.  AND we do NOT use a microwave for Bouins step, everything
was done the old fashioned long way. It you microwave Bouins, the
postfixation may not be adequate (this came from Jerry Fredenburgh - RA
guru also with the staining kits!) and this can affect your staining

At 11:25 AM 7/12/2004 -0400, you wrote:
>Hi all!
>I have a question for you liver people out there.. Is there a trichrome 
>stain out there they people use that will stain collagen as usual and NOT 
>stain hepatocytes red or so red like the muscle in the trichrome? I (yet 
>again) have a researcher that would like a trichrome on livers but would 
>like to see the hepatocytes less red or ideally a different color. This is 
>on FFPE tissue. Thanks!
>Kathy Cormier
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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