[Histonet] Masson's Trichrome Troubleshooting

Wed May 30 08:47:39 CDT 2018

Dear Tasha,

well, I think you should modify your protocol:

(1) don’t use microwave
(2) wash in ethanol after Bouin and not in water (70 - 80 %)
(3) tap water as long as Weigert working solution
(4) Bieberich Scarlet is not the correct stain for Masson, you should use Acid fuchsine + Ponceau 2R + Azophloxine
(5) use a higher concentration of phosphortungstic acid, or / and least leave the sections much longer, up to 30 minutes in this step
(6) do not „air dry“, but remove water with ethanol and ethanol with xylene, then you should get very nice results.

Well, and finally, if you allow I would like to offer you the Masson Kit / Masson Goldner Kit our company offers for quite brilliant stainings: 

https://webshop.morphisto.de/catalogsearch/result/?q=Kit+MASSON+trichrom <https://webshop.morphisto.de/catalogsearch/result/?q=Kit+MASSON+trichrom>

Please do not hesitate to contact us, if you have any further questions.

With best regards
Michael

> Am 30.05.2018 um 14:15 schrieb Campbell, Tasha M. via Histonet <histonet at lists.utsouthwestern.edu>:
> 
> Hello all,
> 
> I am having issues with my trichrome stain and I am about to lose my mind!  We just started doing it in house (although at my previous job I had done trichrome by hand for years so I am not a stranger to it. And I never had issues with it).  When I brought it in house at my current lab, I ordered the same kit that I was familiar with.  Its PolySci.  I did the stain about 5 or 6 times and then all the sudden it quit working.  There was red where it should be blue.  And there was blue staining but it was all in the crypts.   I tried tweaking the stain a few times and nothing worked so I got a new kit.  The first 2 times I used the new kit, it worked perfect!  But after that it is back to doing the same thing again!  The collagen is not staining blue.  It is staining red.  Can anyone please tell me why this is happening?  I never had this issue before!  Thanks in advance!! See my protocol below.
> 
> 1. Mordant in Bouin's solution, microwave 5 minutes, allow to stand 15 minutes.
> 2. Wash in running tap water to remove the picric acid, 5 minutes.
> 3. Weigert's Working Hematoxylin, 10 minutes.
> 4. Blue in running tap water for 5 minutes, rinse in distilled water.
> 5. Biebrich scarlet for 5 minutes.
> 6. Rinse in distilled water.
> 7. Phosphotungstic/phosphomolybdic acid for 10 minutes.
> 8. Transfer directly into Aniline blue for 5 minutes.
> 9. Rinse in distilled water.
> 10. 1% Acetic acid for 1 minute.
> 11. Quick rinse and air dry.
> 12. Coverslip
> 
> 
> 
> 
> Tasha Campbell, B.S.,HTL(ASCP)
> Frederick Gastroenterology Associates
> 310 W. 9th St.
> Frederick, MD 21701
> 301-695-6800 ext. 144
> 
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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