[Histonet] Fibrin stain

[Lee & Peggy Wenk]

You can oxidize it with potassium permanganate. It can use used immediately
(maybe a half hour of mixing), but works better if allowed to set overnight.
It will remain good for about a year.

In the School, I like to make up a batch, and have the students oxidize part
of it with the potassium permangante, for use that week. We store this in a
dark bottle, and it's good for about a year. The other part we put in a
clear bottle, and leave on the counter for about 3 months, to allow it to
naturally oxidize. After that, we put it in a dark bottle in the cupboard.
It stays good for about 3 years. So one batch, split, is good for immediate
use and then for longer term use.

PHOSPHOTUNGSTIC ACID - HEMATOXYLIN (PTAH)
20.0 g   Phosphotungstic acid (P2O524Wo3CxH20)
900 mL   Distilled water, hot but not boiling. Heat in microwave.
1.0 g    Hematoxylin (CI 75290)
10.0 mL  Absolute ethanol, reagent
Dissolve together phosphotungstic acid in HOT distilled water. Allow
phosphotungstic acid solution to cool. Dissolve together hematoxylin in
absolute alcohol. Mix together hematoxylin solution and cooled
phosphotungstic solution. The solution ripens naturally in several (3-4)
months if allowed to set on the counter in light in a clear bottle. After
that, transfer to a dark bottle, and store in the cupboard. Will be stable
for at least one year, up to about 3 years. Store at room temperature. Do
NOT reuse. (If the stain is needed for immediate use, add 0.177 g potassium
permanganate to the solution. When artificially ripened solution is a
reddish-brown color, it is ready for use.)

To get around the Zenker post-mordant, we found a procedure in Ann Preece
that said to post-mordant the slide in Bouins at 60 degree C. for 1 hour.
This works much better than the Zenker (more even staining), and we don't
have to worry about mercury.

Just follow the rest of the procedure after post-mordanting.

Peggy A. Wenk, HTL(ASCP)SLS
Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bobrowitz,
Carol
Sent: Monday, November 17, 2008 3:45 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Fibrin stain

Hello Everyone,

 

I need to do some staining for fibrin.

One stain will be the PTAH.

I would make up the stain but ripening time I don't have.

Any recommendations for a vendor who makes and supplies the PTAH stain
solution?

 

Also an acid red 35 stain has been mentioned.  

I'm pretty sure I've been told the wrong acid red.

There are stains for fibrin that use an acid red.

Any recommendations would be appreciated?

 

Thank you in advance for your help.

 

Carol Ann Bobrowitz HT/HTL

Department of Physiology

Medical College of Wisconsin

414-456-8179

 

cbobrowi <@t> mcw.edu

 

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