[Histonet] carstairs stain

[Monfils, Paul]

CARSTAIR'S METHOD FOR FIBRIN AND PLATELETS

FIXATION:  Formol-saline for at least 48 hours or longer.  (Regular buffered
formalin works fine, but fixation time is important - see results)

Cut paraffin sections at 5 microns.

SOLUTIONS:

5% aqueous ferric alum

Mayer's Hematoxylin

Picric Acid-Orange G Solution:
Saturated aqueous picric acid ................ 20 ml
Saturated picric acid in isopropanol ........ 80 ml
Orange G ............................................. 0.2 gram

Ponceau-fuchsin Solution:
Distilled Water ...................................... 100 ml
Acetic Acid .............................................. 1 ml
Acid Fuchsin ..........................................0.5 gram
Ponceau 2R ...........................................0.5 gram

Aniline Blue Solution:
1% Acetic Acid ..................................... 100 ml
Aniline Blue ............................................ 1.0 gram

1% aqueous phosphotungstic acid solution

TECHNIQUE:

1. Deparaffibnize and hydrate sections

2. Mordant in 5% ferric alum, 5 minutes.  Rinse in running trap water.

3. Stain with Mayer's Hematoxylin, 5 minutes.  Rinse in running tap water.

4. Stain in Picric Acid-Orange G solution, 30 minutes to 1 hour. Rinse once
in distilled water.

5. Stain in Ponceau-Fuchsin solution, 2 to 4 minutes.

6. Differentiate in 1% phosphotungstic acid solution until muscle is red and
background is pale pink. Rinse in distilled water.

7. Stain in Aniline Blue solution, 1 hour.  Rinse in several changes of
distilled water.

8. Dehydrate, clear, coverslip using a synthetic mounting medium.

RESULTS:
Fibrin: (fixation  48 hours or more) - bright red; (less than 48 hours) -
orange to red-orange
Platelets: (fixation 48 hours or more) - gray-blue to navy blue; (less than
48 hours) - light gray
Collagen - bright blue
Muscle - red
Red Blood Cells: (fixation 48 hours or more) - clear yellow; (less than 48
hours) - variable - red, green, or yellow.




> ----------
> From: 	histonet-bounces <@t> lists.utsouthwestern.edu on behalf of
> holling <@t> med-in.uni-saarland.de
> Sent: 	Monday, July 25, 2005 12:45 AM
> To: 	histonet <@t> lists.utsouthwestern.edu
> Subject: 	[Histonet] carstairs stain
> 
> Hi,
> 
> I would be grateful if somebody could give me the carstair stain
> procedure.
> 
> TC Nicole Hollinger
> 
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