RE: [Histonet] Question on Von Kossa

From:"Patsy Ruegg"



That is how I do VK, I put it in the window for 20 min or so.  I do a
special H&E counterstain on mine, using an aqueous eosin.  The eosin will
light up the osteoid by fluorescing under uv light.  We use this with a
image analysis system to measure total area, calcified bone area (light
scope, from the black silver stain) osteoid seam thickness (fluorescent
scope using the eosin fluorescent property, everything else will be dark
except the osteoid seams) if you labeled the bone with a fluorchrome you can
just look at an unstained section to measure area between two labels.
Patsy

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Friday, October 17, 2008 10:15 PM
To: Herrick.James@mayo.edu; histonet@lists.utsouthwestern.edu
Subject: [Histonet] Question on Von Kossa

James,
     Try this: once the sections are brought to water, 5% silver nitrate in
either bright sunlight or a 60-100 watt incandescent light bulb for 30 min
(check for browning of the control tissue adjust time as needed). Rinse well
in distilled water. 5% Sodium Thiosulfate for 1 min. Rinse and counterstain
with nuclear fast red or whatever you think would look cool :-). Dehydrate,
clear and coverslip the slides.
   Of course, as with any silver stain use acid cleaned glassware and gloves
unless you like watching your fingers turn black in the sunlight. Really
well decalcified tissue usually has the Calcium washed out. Try a calcified
breast or something that wasn't decalcified. This causes microtomists to
curse a lot, but it makes great VonKossa controls.

Have fun,
Amos

Message: 10
Date: Thu, 16 Oct 2008 15:45:11 -0500
From: "Herrick, James L." 
Subject: [Histonet] Question on Von Kossa
To: 
Message-ID:
       
Content-Type: text/plain;       charset="iso-8859-1"

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 Ám thick)? I would appreciate it
greatly. Thank you much.

Jim
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