[Histonet] VonKossa's calcium stain. Counterstain etc

[John Kiernan]

Any haemalum will do, but blue is not the most pleasing contrast to go with the black silver deposits. Pink or light red is better. Here are three pink to red counterstains, all traditional. Check in a textbook or manual for instructions.
 
   1. Neutral red (CI 50040) is good: 0.5% in water; adjust to pH4 with acetic acid; stain for about 2 minutes; the solution keeps for at least 5 years and can be used repeatedly.  
   2. Safranine O (CI 50240) can be used similarly but needs a longer staining time. 
   3. Nuclear fast red (CI 60760) is also OK: NFR 0.2G, aluminium sulphate crystals 10G, water 200ml; heat until it boils, cool overnight, decant and filter; stain for 5-10min. The solution is good for about a year; always filter before using.
 
The Biological Stain Commission has standards for certification of all three of these dyes. Certified neutral red and safranine O have been available for many (50+) years. Nuclear fast red was only recently added to the BSC's list. See Frank et al 2007. Certification procedures for nuclear fast red (Kernechtrot), C.I. 60760. Biotechnic & Histochemistry 82: 35-39. Certified NFR powder may not yet be available to labs or to vendors of stain solutions. If you buy a ready-made solution of any dye you should choose one that was made from a BSC-certified batch of the powder. 
 
To any vendors of dye powders who read this message: email me for more information about the certification criteria for nuclear fast red.  Also, check out http://www.biologicalstaincommission.org and click on one of the "Vendors" tabs or links.
 
John Kiernan
Anatomy,  UWO
London, Canada
= = =
----- Original Message -----
From: karine cadoret <kcadoret <@t> amc.edu.au>
Date: Thursday, July 16, 2009 22:36
Subject: [Histonet] VonKossa's calcium stain
To: histonet <@t> lists.utsouthwestern.edu

> Hi, 
> 
> When doing a VonKossa stain in order to demonstrate calcium in tissue,
> does it matter much if I use Mayer's hematoxylin instead of Ehrlich's
> hematoxylin (which takes 6 months to ripen) ? 
> 
> Also, can I simply use homemade scott's tapwater for blueing 
> instead of
> using a lithium carbonate solution ? 
> 
>  
> 
> Thank you for your help, 
> 
>  
> 
> Karine Cadoret
> 
> Fish health laboratory manager
> 
> National Center for Marine Conservation and Resource Sustainability
> 
> Newnham, TAS
> 
> Australia 
> 
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