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 < [email protected]> 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:[email protected]] > > Sent: Tuesday, September 10, 2019 8:43 AM > To: '[email protected]'; 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 <[email protected]> > Sent: 09 September 2019 10:53 > To: '[email protected]' <[email protected] > > > 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: [email protected] > texasheart.org< > https://urldefense.proofpoint.com/v2/url?u=https-3A__www.texasheart.org_&d=DwICAg&c=iORugZls2LlYyCAZRB3XLg&r=7cy9qXFa73jDX2Iixpjkq1XlWAfHgLLHm33agI_sCKA&m=gOhsr6cogN3TBZh8C2dHLiczk4rN5grOBAD1YkzJlrk&s=8BI3BsDaT4RpUA7gRKKiLai8Lsjj8dijlm_QRu3kElg&e= > > | facebook< > https://urldefense.proofpoint.com/v2/url?u=https-3A__www.facebook.com_Texas.Heart.Institute&d=DwICAg&c=iORugZls2LlYyCAZRB3XLg&r=7cy9qXFa73jDX2Iixpjkq1XlWAfHgLLHm33agI_sCKA&m=gOhsr6cogN3TBZh8C2dHLiczk4rN5grOBAD1YkzJlrk&s=deGJye4k3LKtsaLIxFu7_k3FNXU5R6uqcgIRXry7oug&e= > > | twitter< > https://urldefense.proofpoint.com/v2/url?u=https-3A__twitter.com_Texas-5FHeart&d=DwICAg&c=iORugZls2LlYyCAZRB3XLg&r=7cy9qXFa73jDX2Iixpjkq1XlWAfHgLLHm33agI_sCKA&m=gOhsr6cogN3TBZh8C2dHLiczk4rN5grOBAD1YkzJlrk&s=r7bgnhb3v6jPkFz2Km9Nf_p5ltR1MFAjWKurvn82-GI&e= > > > _______________________________________________ > Histonet mailing list > [email protected] > > https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu_mailman_listinfo_histonet&d=DwICAg&c=iORugZls2LlYyCAZRB3XLg&r=7cy9qXFa73jDX2Iixpjkq1XlWAfHgLLHm33agI_sCKA&m=gOhsr6cogN3TBZh8C2dHLiczk4rN5grOBAD1YkzJlrk&s=Tg-ltd7S0Gvp2_-XSvBjsRuw1esVa8scKHMk6uYj10c&e= > _______________________________________________ > Histonet mailing list > [email protected] > > https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.utsouthwestern.edu_mailman_listinfo_histonet&d=DwICAg&c=iORugZls2LlYyCAZRB3XLg&r=7cy9qXFa73jDX2Iixpjkq1XlWAfHgLLHm33agI_sCKA&m=gOhsr6cogN3TBZh8C2dHLiczk4rN5grOBAD1YkzJlrk&s=Tg-ltd7S0Gvp2_-XSvBjsRuw1esVa8scKHMk6uYj10c&e= > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- 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 [email protected] <[email protected]>* _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
