Jim, Here you go: MMA Thin (5 microns) Sections 1) Deplastify with three changes of Xylenes @ 60C for 30-60 minutes (depending on size of section). You can also use three changes of acetone @ RT for 15-30 minutes. 2) Hydrate tissue from ethanol to DI water (100% EtOH for 5 min, 95% EtOH for 5 min, 70% EtOH for 5 min, DI H2O for 5 min) @ RT. 3) Stain in 5% Silver Nitrate solution @ RT for 5 minutes. Keep tissue and solution protected from light!!!! 4) Three DI water rinses @ RT for one minute each. 5) Develop all bound silver ions in Sodium Carbonate-Formaldehyde solution (5 g sodium carbonate, 25 mL formaldehyde, 75 mL DI water) @ RT for 2 minutes (TIME CRITICAL). 6) Two DI water rinses @ RT for one minute each. 7) Stop reaction and remove all unbound silver ions in Farmer's Diminisher (20 g sodium thiosulfate, 1 g potassium ferricyanide, 210 mL DI water) @ RT for 30 seconds. This solution is only stable for 30-45 minutes so make and use fresh. 8) Wash in running tap water for 20 minutes. 9) Rinse in DI water @ RT for one minute. 10) Counterstain with 5% MacNeal's tetrachrome solution. There are other choices, but this is what I use. 11) Dehydrate to Xylenes and coverslip. I will send you a copy of my detailed protocol. Jack
> Date: Thu, 16 Oct 2008 15:45:11 -0500> From: [EMAIL PROTECTED]> To: > histonet@lists.utsouthwestern.edu> Subject: [Histonet] Question on Von Kossa> > > Hello everyone,> > Does anyone have a good Von Kossa stain protocol that > they would not mind sharing, on animal bone tissue (femur/tibia) that has > been embedded in GMA or MMA (sections are between 5 and 10 µm thick)? I would > appreciate it greatly. Thank you much.> > Jim> > > > > _______________________________________________> Histonet mailing list> > Histonet@lists.utsouthwestern.edu> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Want to read Hotmail messages in Outlook? The Wordsmiths show you how. http://windowslive.com/connect/post/wedowindowslive.spaces.live.com-Blog-cns!20EE04FBC541789!167.entry?ocid=TXT_TAGLM_WL_hotmail_092008_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet