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

[Anna Huntley Coffey]

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