[Histonet] Histonet mailing list submission: Jones' Methenamine Silver Stain

[Tony Henwood (SCHN)]

Anna,

I would try Hayashi et al (Stain Technol 64(4):185-190, 1989) technique. They have modified this procedure by including a treatment with thiosemicarbazide before silver impregnation. This enhances the deposition of silver, decreasing background staining. 
It is believed that the hydrazine group (-NHNH2) of thiosemicarbazide blocks the aldehyde group produced following periodic aid treatment and the thiocarbamyl group (-SCNH2) of thiosemicarbazide reduces the methenamine silver.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Anna Huntley Coffey
Sent: Tuesday, 16 July 2013 7:13 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Histonet mailing list submission: Jones' Methenamine Silver Stain

Dear Histonetters,

I was wondering if anyone has experience with Jones' Methenamine Silver stain?  We have attempted this stain several times, each time with a slight tweak to the protocols we found online.  We found that it took much longer than expected (3-4 hours compared to the expected 1 hour) for the membranes to develop the expected black color every attempt we made.  The few times we did see the black color develop in the membranes, the next step in the gold chloride solution (1 minute) washed away all the dark color in the kidney, leaving it a very light brown.  However, we did notice that only the skin sample on our test slides retained the black color.  Since we ultimately need this stain for kidney samples, this didn't do us any good.
 We tried thinner sections (2 micron), acid washed glassware, brand new reagents, and multiple protocols with no luck.  I would greatly appreciate any advice as to what we could try differently or if anyone else has experienced these same problems.

Thank you!
Anna
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