[Histonet] Mallory-Azan stain

Wed Sep 11 13:42:14 CDT 2019

Thank you John,
As always,  a wealth of information. I did do a PAS for him, I am not totally sure he knows exactly what he is looking for.
I always learn something from your posts.
Betsy

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 <www.texasheart.org> | facebook <www.facebook.com/Texas.Heart.Institute> | twitter <twitter.com/Texas_Heart>
-----Original Message-----
From: Colleen Forster via Histonet <histonet at lists.utsouthwestern.edu> 
Sent: Tuesday, September 10, 2019 11:17 AM
To: Morken, Timothy <Timothy.Morken at ucsf.edu>
Cc: Histonet <histonet at lists.utsouthwestern.edu>; John Kiernan <jkiernan at uwo.ca>
Subject: Re: [Histonet] Mallory-Azan stain

*** Important*** This email is not from Texas Heart Institute. Only click links or open attachments you know are safe.
________________________________

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
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--
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
B173 PWB      612-626-1930

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