Re: stain for platelet & fibrin

From:Renee Escalona <rescalona@tvmdl.tamu.edu>

Cathy,

Our lab uses Carstairs Method for fibrin and platelets.  Here it is:


Fixation:  NBF  (formalin saline recommended)

Sections: 5 microns

Solutions:

Picric acid-Orange G Solution:       20 ml  saturated picric acid aqueous
                                                      80 ml   saturated picric acid in isopropanol
                                                       0.2 gm  Orange G

Ponceau-Fuchsin Solution:        0.5 gm   acid fuchsin
                                                  0.5 gm    Ponceau 2R
                                                   1 ml   glacial acetic acid
                                                   99 ml   distilled water

Aniline Blue Solution:            1 gm   aniline blue
                                             100 ml   1% acetic acid


Procedure:

1.   Deparaffinize and hydrate to water.
2.   Mordant in 5% ferric alum for 5 minutes and then rinse in tap water.
3.   Stain with Mayer's Hematoxylin for 5 minutes and rinse in tap water.
4.   Stain in picric acid-Orange G solution for 30 minutes to 1  hour.
5.   Rinse once in distilled water.
6.   Differentiate with 1% phosphotungstic acid.
7.   Stain with aniline blue solution for 30 minutes to 2 hours.
8.   Rinse in several changes of distilled water.
9.   Dehydrate, clear and mount. 

Results:

fibrin - bright red                                        muscle - red
platelets - gray-blue to navy                      RBC's - clear yellow
collagen - bright blue


from Theory & Practice of Histotechnology, Second Edition, page 281


Good luck Cathy!

Renee Escalona, BS, HTL(ASCP)
Laboratory Supervisor
Texas Veterinary Medical Diagnostic Lab
College Station, TX

>>> Cathy Mayton <Cathy@wasatchhisto.com> 06/28/01 02:23PM >>>
Fellow Histonetters,

We are looking for stain specific for platelets and fibrin.  The tissues
are iliac or femoral arteries that will be embedded in paraffin.  Is the a
special stain specific to platelets and fibrin or is there an IHC method
that would be more specific?

Thanks in advance,
Cathy






*********************************
GLP Compliant Laboratory

Cathy A. Mayton
Project Director
Wasatch Histo Consultants, Inc.
www.wasatchhisto.com 






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