Thanks for the reply Toysha. That was a typo. I should have typed 0.5%.
Thanks again. Hope all is well!
Betsy
Betsy Molinari, HT (ASCP)
Sr. Histology Research Technician
CV Pathology Research
Texas Heart Institute
6770 Bertner Avenue, MC 1-283
Houston, TX 77030
Office: 832-355-6524 | Fax: 832-355-6812
Email: bmolin...@texasheart.org
texasheart.org | texasheartmedical.org
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-Original Message-
From: Mayer,Toysha N via Histonet
Sent: Wednesday, May 19, 2021 1:04 PM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Movats
*** Important *** This email is not from Texas Heart Institute. Only click
links or open attachments you know are safe.
Hey Betsy,
It could also be the woodstain scarlet and the saffron. I would omit the 5%
acetic after the phosphotungstic.
Take care,
Toysha Mayer
Message: 1
Date: Fri, 7 May 2021 22:11:30 +
From: John Kiernan
To: "histonet@lists.utsouthwestern.edu"
,Betsy Molinari
Subject: Re: [Histonet] Movats
Message-ID:
Content-Type: text/plain; charset="iso-8859-1"
Dear Betsy,
Don't say you are sorry for putting a long post on Histonet! To get
troubleshooting help you need to say exactly what you did. If you wrote only,
"why are my sections brown after Movat staining", nobody would understand your
problem.
Your procedure starts with an hour in hot Bouin. For many years this has been a
routine prior to trichrome stains done on sections of specimens fixed in
neutral formaldehyde. It isn't part of Movat's original method (Arch. Path.
60:209-295, 1955), which probably was devised for sections optimally fixed for
trichrome staining (in mixtures containing mercuric chloride).
Movat's pentachrome is a trichrome method preceded by alcian blue (for no
obvious reason) and an iron-haematoxylin for nuclei and elastin. It differs
from older trichromes in using a mixture of yellow polyene dyes (saffron) to
stain the collagen, instead of the blues or greens as in the Mallory and Masson
methods.
Your method includes "5% sodium thiosulfate -1 min" after the iron-haematoxylin
stain for black nuclei and elastic fibres. This also isn't part of Movat's
pentachrome method, and I wonder why. Did you inherit an informal list of
instructions passed on within the lab? After a mercuric fixative, hydrated
sections are dipped in iodine, followed by thiosulphate, before staining, to
remove a black deposit (probably mercurous chloride) introduced by the
fixative.I've been seeing similar informal passing of bad staining instructions
in research labs for many years. Are you a victim of this trend?
The thiosulphate step in your procedure obviously does no harm, because you got
the right results with the dog tissues. There may be something different about
your human specimens: perhaps inadequate fixation, or excessive acid treatment
(if that's what Cal rite is) for decalcification.
If the sections of human arteries look OK with a microscope, it might not
matter that grossly they are a different colour from the dog small intestine
sections. They are, after all, different tissues.
A rather long, and not very helpful reply!
John Kiernan
London, Canada
= = =
From: Betsy Molinari via Histonet
Sent: May 5, 2021 9:46 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Movats
Hi Histonetters,
I have received several human vessels for paraffin processing and to stain the
sections for H&E and Movats. The H&E were fine. The human sections turned
brownish yellow with the Movats.The control which is canine small intestine was
perfect.
The protocol is standard
Bouins 1hr in 58C waterbath
Rinse till yellow disappears
Rinse in DH2O
1% Alcian Blue -20 min
Rinse in running tap H2O -5min
Alkaline alcohol-1hr
Rinse 10 min tap H2O
Rinse in DH2O
Verhoff's Hematoxylin -15 min
3 changes DH2O
Differentiate in 2% FeCl
Rinse in DH2O
5% sodium Thiosulfate -1min
Rinse in running tap-10 min
Rinse in DH2O
Woodstain scarlet/acid fuchsin-1.5 min
Rinse in DH2O
Rinse in 0.5% acetic acid water
5% aqueous phosphotungstic acid -2 changes 5 min each Rinse in 5% acetic acid
water Rinse in 3 changes absolute ETOH 6% alcoholic Safran solution Absolute
alcohol-xylene-coverslip The human slides were fine until the Safran step. When
I removed them from the stain into the 100% they were a yellowish brown .Under
the scope the colors were there, blue, red, yellow and black. But on the slide
the tissue was that brownish yellow. The researcher does not like to strong
yellow color. Since my control was fine I question if something was going on
with their tissue. I do not know how the tissue was handled before it came into
the lab. They were very calcified and were decaled for 1-3 days in Cal Rite. I
do know they were not rinsed after decal and were put straight back into 10%
NBF before I got them for processing.
Should I have used a human control inste
