[Histonet] Herlant Pituitary Stain II

John Kiernan jkiernan <@t> uwo.ca
Tue Aug 21 12:07:23 CDT 2007

This method is a simplification of Adams's pituitary stain, which is the sequence: 
Performic or peracetic acid - oxidizes cystine -S-S- to cysteic acid, a sulphonic (strong) acid.
Alcian blue pH 0.2  - stains the generated sulphonic acid groups (TSH cells and neurosecretory material of the posterior lobe).
PAS - stains neutral glycoproteins, especially cells producing gonadotrophic hormones
Orange G in PTA - stains growth hormone cells.
Mayer's Haemalum - for nuclei.

I've not read Herlant's 1960 paper, the one cited in Humason's Animal tissue Techniques, but have read a related one: Pasteels & Herlant 1962 Z. Zellforsch. 56: 20-39. The principal innovation was replacing performic or peracetic acid with acidified potassium permanganate followed by a bisulphite rinse to remove brown MnO2 from the sections. You can use 1% oxalic acid instead of bisulphite.
I've done these methods and recommend permanganate over the organic per-acids, which are really noxious. Some histochemists have argued against permanganate oxidation (see books by A.G.E.Pearse and by M.Gabe) but none can deny that permanganate-H2SO4 followed by alcian blue pH 0.2 gives good staining of neurosecretory material. Mast cells are also blue because of their heparin content, but that staining does not require the prior oxidation. 

In the late 1950s rational techniques of this kind replaced earlier staining techniques for pituitary cell-types, which were mostly variants of Heidenhain's AZAN and other trichromes. Since the 1970s antibodies to all the anterior and posterior pituitary hormones have been available and the cell types can be unequivocally identified by immunohistochemistry. 

John Kiernan
Anatomy, UWO
London, Canada
----- Original Message -----
From: Andrea Grantham <algranth <@t> u.arizona.edu>
Date: Tuesday, August 21, 2007 11:31
Subject: [Histonet] Herlant Pituitary Stain II
To: histonet <@t> lists.utsouthwestern.edu

> Good Morning!
> I am getting ready to do a Herlant Stain on pituitary and I am 
> confused by the protocol in Humason's Animal Techniques, 3rd edition.
> If there is anybody out in histoland who is familiar with this 
> stain 
> OR if you have this book and can help with interpretation of the 
> protocol please let me hear from you.
> Thanks,
> Andi Grantham
> .....................................................................
> : Andrea Grantham, HT(ASCP)     Dept. of 
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