[Histonet] Verhoeff/Masson's Stain

Tony Henwood AnthonyH <@t> chw.edu.au
Tue Mar 22 18:23:04 CDT 2011

See there are always differing experiences (see my earlier post)

How are you Bryan, well I hope?

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn
Sent: Wednesday, 23 March 2011 5:48 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Verhoeff/Masson's Stain

Verhoeff's staining solution uses an iron mordasnt, which is an oxidising agent, and iodine, another oxidising agent.  It is very easy to overoxidise the hematoxylin, particularly if a stock alcoholic solution is used.  I always used to preweigh 1 gram hematoxylin and keep in test tubes, then add the required amount of ethanol at the time I made the working solution.  Problems I had had with poor staining then stopped.

Have you considered switching to an iron resorcin dye type method?  They are progressive, much easier to control and work very consistently. 
Their drawback is that making the solution is a nuisance and staining often needs overnight. I liked Humberstone's variant (greeny blue black
elastic) as the solution is stable for several years and improves over that time with shorter staining times.  Others like Miller's (black elatic).  There are others as well.

Bryan Llewellyn

John Shelley wrote:
> Hello Histonetters,
> I have been asked to work on a project that will require me to do a Verhoeff/Masson's stain. I have run some samples through the stain from a protocol that I found on the archives and it is not working to my satisfaction. I am not getting the nice elastin fibers to show black like they should. I have looked at the slides just before going into 2% ferric chloride and it appears to be dull or limited staining. I am using a kit from PolyScientific and not sure if that might be my problem or not. My question is, are there another people who are running this special stain combo and would you be willing to share the procedure and also who you are buying your reagents from to accomplish the staining. I appreciate any help with this request. Thanks!!!
> Kind Regards!
> John J Shelley
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