Re: pituitary staining

From:Lee & Peggy Wenk <>

Pituitary cells in the adenohypophysis part (hormone secreting anterior
portion) can be divided into acidophils, basophils and chromophobes, in
terms of histology histochemical staining.

Acidophils are the somatotrophs (growth hormone) and lactotrophs
(prolactin). These both stain with acid dyes, such as eosin, phloxine,
orange G and some of the red dyes used in trichromes.

Basophils are the corticotrophs (adrenocortical stimulating hormone ACTH),
thyrotrophs (thyroid stimulating hormone TSH), and gonadotrophs (follicle
stimulating hormone FSH or leutenizing hormone LH). These stain well with
basophilic dyes such as hematoxylin, aniline blue, methyl blue, and even
aldehyde fuchsin, if I remember correctly. They also stain with periodic
acid-Schiff (PAS) since they have glycogen compound on the protein hormones.

So if you want to differentiate acidophils from basophils, use a
combination, such as a trichrome stain (Masson or Mallory or Montreal used
to be favorites for these) or MSB (eg. aniline blue - brilliant crystal
scarlet) or a PAS-orange G.

I know the John Bancroft histology book (Theory and Practice of Histological
Techniques) has these procedures. Some procedures are on the web page from
his hospital in Nottingham, England (put on web by Jim Lowe). The only way I
know to get there is to go to the Histotech's Home Page:

click on  Peggy's links
click on Stains and Tissues
click on Histopathology Laboratory Methods, and go to procedures

Another source would be Brian Llewellyn from Canada, who also has some
procedures for pituitary on his web site. Again, the only way I know to get
there is through the Histotech's Home Page:

click on Peggy's links
click on Stains and Tissues
click on Stains File, then go to Procedures, and either look for pituitary,
or look for the names of the staining procedures.

However, a word of warning about histology stains. The majority of cells in
the pituitary and pituitary adenomas are not acidophils or basophils - they
are chromophobes. In other words, they do not stain with acid or basic dyes.
They may either be the precursors of the cells, or cells in various stages
of degranulation. So they may be acidophils or basophils that we cannot
stain at this point of their development.

Yet, when immunohistochemistry (IHC) is performed on these chromophobes,
many WILL stain positive for their hormone. However, not all will stain with
immunohistochemistry. These chromophobes may have totally lost their

But currently, I would say the majority of labs are using IHC, supplemented
by electron microscopy study of the granules, to differentiate pituitary
cell types. Very few labs will use histochemical staining any more.

Hope this helps with your research project.

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

----- Original Message -----
From: "carlos ballestas" <>
To: <>
Sent: Thursday, April 19, 2001 11:02 AM
Subject: pituitary staining

> hello my name is carlos ballestas, I am a research tech at the
> university of south florida and am currently working with pituitary
> cells. I wanted to ask if you knew of a staining method for pituitary
> cells to distinguish the somatatrope and the lactatropes form the rest
> of the cells. If so I would apreciate it if you would let me know.
> thank you very much for your time and cooperation
> sincerely
> carlos ballestas

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