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

Rena Fail renafail2 <@t> gmail.com
Mon Jul 15 18:06:53 CDT 2013

place your coplin jar with silver solution in a 65 degree waterbath    5-10
minutes before your slides need to go in the sol. PreHeating the sol.in the
waterbath  and staining the slides in the waterbath will aid in reducing
the time down to 10-15 minutes. make the methenamine- silver fresh. the
silver and borax can be made ahead the methenamine should be made fresh
each time.,    Don;t check the slides until the tissue looks golden
brown.your end point is when the elsatic tissue is black (wlastic stains1st
with the silver), Bowman's capsule is black and the basement membrane of
the glomerlus is golden brown to black. the temptation is to stop when the
bm is defined. Make sure the bowmans capsule is black and the bm in the
glom. is dark golden brown to black. BM in thhte skin stains rapidly much
faster than thhat in thhe glom

Rena fail

On Mon, Jul 15, 2013 at 5:13 PM, Anna Huntley Coffey <annamhuntley <@t> gmail.com
> wrote:

> 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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