Paul,
For MMA embedded specimens (MMA +DBP), I first deplastify my sections, hydrate
to water, stain in 5% silver nitrate solution for 5 minutes (in the dark), wash
times three changes in DI H2O (in the dark), develop in sodium-carbonate
formaldehyde solution for 2 minutes (in the dark), wash times two in DI H2O
(back under normal lighting conditions), then stop the reaction in sodium
thiosulfate + potassium ferricyanide solution for 30 seconds, and immediately
rinse in running tap water for 15 minutes.
The Von Kossa reaction results from process above then yields black mineralized
bone. After the tap water rinse, I generally counterstain with 2% MacNeal's
tetrachrome for 5 minutes, rinse in DI H2O, and dehydrate to xylenes to
coverslip. This then reveals immature bone formation or osteoid = grayish or
jaded green, growth plate cartilage = purple, osteoblasts = blue, osteoclasts =
blue-green, bloods cells = greenish, etc.
Feel free to contact me if you have any additional questions.
Jack
Date: Thu, 12 Mar 2009 11:19:49 -0400
From: pmonf...@lifespan.org
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Von Kossa staining on PMMA sections
The standard Von Kossa silver stain for calcium calls for 20 minutes in the
silver nitrate solution under UV light. There is a modified Von Kossa for
plastic embedded bone sections, which is identical except it calls for a
minimum of 6 hours in the silver nitrate solution under UV. Does anyone know
why such a long staining time is recommended? Visually the calcium in the
bone sections turns black within 20 minutes, so why is so much additional
time needed? Thanks.
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