RE: [Histonet] (no subject)

From:"Bernice Frederick"

I have a researcher that likes me to do the trichrome over the EVG. It's a
combo of both stains. Looks gorgeous. I use an aorta as the control when I
do it.She also has me do Goldner's trichrome on occasion. Looks good on
heart muscle.

Bernice Frederick HTL (ASCP)
Northwestern University
Pathology Core Facility
710 N Fairbanks Court
Olson 8-421
Chicago,IL 60611

-----Original Message-----
[] On Behalf Of Tony Henwood
Sent: Thursday, July 03, 2008 6:02 PM
To: Sheree Beam;
Subject: RE: [Histonet] (no subject)


Masson's stain is quite capricious.

Unlike the van Gieson stains, over differentiation of the red muscle
stain is common. The use of mordants can often help.
Try placing hydrated slides for 30min in Bouins solution (or aqueous
saturated picric acid) at 60oC prior to staining.
With the use of the blue collagen counterstain, confusion can also occur
since under-differentiation of the nuclear stain, Celestine
blue-Harris's Hx or some variant of the iron haematoxylin, can render
the muscle blue. The blue you are describing could be due to
overstaining with the celestine blue-Hx rather than the iron Hx which
would look black. Again overstaining with the blue dye of the Massons as
well as overdifferentiation of the red dye would render muscle blue.

In my experience control sections rarely behave like diagnostic slides
with the Massons. Different fixation times and older control sections
change in their affinity for the red and blue stins of the Massons.

Try a green collagen stain rather than the blue. The photos may look
more pleasing or have you considered using a Van Gieson's stain in


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318

-----Original Message-----
[] On Behalf Of Sheree
Sent: Friday, 4 July 2008 5:28 AM
Subject: [Histonet] (no subject)



I received Masson's trichrome stains of arteries and adjacent muscle
from a histolab.  The first time I received them, the entire artery was
blue and the adjacent muscle was purple to sometimes red.  The smooth
muscle in the arteries were completely blue.  However the control tissue
of kidney which included a small artery stained perfectly.  I asked for
the tissue to be recut and try again with Masson's.  This time the
smooth muscle in the artery has some magenta, but in other areas the
blue prevailed.  More controls were run and all were perfect.  The
tissues were fixed in 10% neutral buffered formalin for 10 or 30 days as
there were 2 groups.  We used two different boxes of formalin.  After
trim, the tissues were placed back into formalin and sent to the histo
lab.  There was no fixation in ethanol.


Are there any explanations out there?  Is there anything we can do?  I
need the trichromes for photomicrographs.  Would a different trichrome
stain potentially help?  All input would be appreciated.


Thank you,




Sheree Beam, DVM, MS, ACVP, Diplomate

Veterinary Pathologist

Preclinical Pathology Consulting Service, LLC


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