Anna, 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 <annamhunt...@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 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet