--On 05 June 2001 11:26 -0400 Jill Songer <jtsonger@vt.edu> wrote:

> Hi Netters,
>
> One of our pathologists has requested stain for hemoglobin. Okajima
> perhaps?? If someone could send me this procedure, I will be forever
> grateful.
>
> **************************************************************************
> Jill Songer HT (ASCP)
> Supervisor, Anatomic Pathology
> Virginia Tech
> Veterinary Medical Teaching Hospital

Dear Jill,

The following technique is from 'Histopathologic Technic and Practical 
Histochemistry' Third edition by Lillie.

The Okajima Stain [Anat. Rec. 11:295, 1916] - Dunn's modification.

Fix in 10% neutral buffered formalin.

1. Take sections to water.
2. Mordant for 1 minute in 10% phosphomolybdic acid.
3. Wash in distilled water.
4. Stain for 1 hour in 10% phosphomolybdic acid 9ml, saturated aqueous 
[7.69%] alizarin red S [C.I. No 58005] 30ml.
[Okajima; 20 minutes to 20 hours].
5. Wash in distilled water.
6. Counterstain if desired with an unacidified alum haematoxylin for 3-5 
minutes.
7. Wash in water, dehydrate, clear and mount in D.P.X.

Results:- Haemaglobin is light to dark orange red.
The background is light brown with haematoxylin, orange without 
haematoxylin.

Hope this helps,

Tony Brandwood,
Dept. of Veterinary Pathology,
University of Liverpool,
P.O. Box 147,
Liverpool,
L69 3BX.
0151 794 425


Best wishes,





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