[Histonet] Mallory-Azan stain

Tue Sep 10 11:17:27 CDT 2019

Wow John,

Great information.,....always so much to learn!~

Thank you,

Colleen Forster
U of MN

On Tue, Sep 10, 2019 at 10:55 AM Morken, Timothy via Histonet <
histonet at lists.utsouthwestern.edu> wrote:

> John, we love it when you "ramble!" It gives us an appreciation for the
> history and breadth of histo techniques.
>
> Tim Morken
> Supervisor, Electron Microscopy/Neuromuscular Special Studies
> Department of Pathology
> UC San Francisco Medical Center
>
> -----Original Message-----
> From: John Kiernan via Histonet [mailto:histonet at lists.utsouthwestern.edu]
>
> Sent: Tuesday, September 10, 2019 8:43 AM
> To: 'Histonet at lists.utsouthwestern.edu'; Betsy Molinari
> Subject: Re: [Histonet] Mallory-Azan stain
>
> Mallory's (easy) and AZAN staining (difficult) are different methods!
>
> Frank B. Mallory's trichrome stain  (Journal of Medical Research 13:
> 113-136, 1905) is the earliest and one of the simplest of its kind: acid
> fuchsine followed by a solution containing orange G, aniline blue and
> phosphotungstic acid (PTA).  Martin Heidenhain's trichrome is usually
> called AZAN (from Azokarmin and Anilinblau, the German names of two of the
> dyes used. I've not read his original 1916 publiication, but a very
> thorough account was given by Manfred Gabe in his 1976 Histological
> Techniques book (ISBN 3540901620), pp. 219-223. I used this quite a bit in
> the 1990s mostly on paraffin sections of Bouin-fixed decalcified rats'
> heads. It is a 15-step procedure taking >2 hours and it includes two
> critical differentiations requiring careful microscopic control.
> Instructions based on my experiences can be found in Histological and
> Histochemical Methods (5th ed., 2015, pp.198-200).
>
> AZAN gives a wider range of colours than Mallory's or Masson's trichrome
> or the various one-step trichromes (Cason, Gomori, Gabe). The related
> Romeis "cresazan" procedure was used to identify at least 6 anterior
> pituitary cell-types until the 1950s when more rational histochemically
> based stains were introduced by Adams, Herlant, Pearse and others.
> Nowadays, immunostainng accurately shows the hormones in pituitary cells,
> but much more expensively.
>
> All trichromes give poor results after simple fixation in neutral
> formaldehyde. Bouin or (better) a mercuric chloride-containing fixative is
> needed. Zinc-formalin is probably also OK. (I haven't tried it myself for
> this purpose). If material fixed in NBF must be used, immerse hydrated
> paraffin sections in saturated aqueous picric acid either for 2h at 56-60C
> or overnight at room temperature, then wash well in water before staining.
> (Bouin's fluid is often used, but its ingredients other than picric acid
> are unnecessary.) Experiments are needed to learn the mechanism of this
> "rescue" of  staining properties of sections formaldehyde-fixed tissue,
> which is sometimes wrongly called "mordanting". My guess is that it's
> comparable to antigen retrieval. It has been claimed that citrate buffer is
> just as good, though the photos are unconvincing (J. Histotechnol. 26, 133).
>
> It should be possible to identify Purkinje fibres with any staining method
> that shows nuclei and myofibrils, such as H&E or a trichrome method simpler
> than AZAN. A glycogen stain such as PAS might show this substance in the
> otherwise pale areas around the central nuclei of Purkinje fibres. I
> suggest persuading your researcher to let you try something simpler before
> attempting Heidenhain's AZAN. Wheater's Functional Histology has a nice
> photomicrograph of a section stained with H&E and for endocardial elastin
> (looks like orcein).
>
> Enough rambling!
>     John Kiernan
> Anatomy & Cell Biology
> University of Western Ontario
> London, Canada
> = = =
>
> ________________________________
> From: Betsy Molinari via Histonet <histonet at lists.utsouthwestern.edu>
> Sent: 09 September 2019 10:53
> To: 'Histonet at lists.utsouthwestern.edu' <Histonet at lists.utsouthwestern.edu
> >
> Subject: [Histonet] Mallory-Azan stain
>
> Hi histonetters,
> I have a researcher that wants to stain Purkinje fibers and has requested
> a Mallory-Azan stain.
> I have no experience with this stain. I have looked online for information
> but am reaching out to you for personal advice.
> Thanks.
> Betsy Molinari HT,ASCP
> Texas Heart Institute
> 6770 Bertner Ave.
> Houston, TX  77030
> 832-355-6524 (lab)
> 832-355-6812 (fax)
>
> Betsy Molinari
> Sr. Histology Research Technician
> CV Pathology Research
>
> Texas Heart Institute
> 6770 Bertner Avenue, MC 1-283
> Houston, TX 77030
>
> Office: 832-355-6524 | Fax: 832-355-6812
> Email: BMolinari at texasheart.org
> texasheart.org<
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-- 
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
B173 PWB      612-626-1930

*If submitting histology request please also forward to Lori Holm at
holml at umn.edu <holml at umn.edu>*