[Histonet] RE: Verhoeff/Masson's Stain

[Tony Henwood]

Hi all,

My experience with Verhoeff stain indicates that poor, muddy staining of elastic fibres results from using too fresh a solution of alcoholic haematoxylin. 
The usual methods require a 10% alcoholic Hx that is mixed with the other ingredients. 
I always let the stock 10% alcoholic Hx stand for at least a week before using. 
Too fresh Hx is also difficult to differentiate (sometimes less than 1 dip is required in 2% ferric chloride).

As for controls, a section of skin is best - the odd artery will stain, but more importantly, the fine elastic fibres in the dermis should be easily seen.

Regards 
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 John Shelley
Sent: Wednesday, 23 March 2011 5:14 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Verhoeff/Masson's Stain

Hi Shirley,

I do make it fresh each time and the reagents are brand new from the company. I am using an appendix as my control and trying to get my hands on some aorta but I really think I should see some elastin fiber staining in the appendix.

Kind Regards!
 
John J Shelley

-----Original Message-----
From: Shirley A. Powell [mailto:POWELL_SA <@t> mercer.edu] 
Sent: Tuesday, March 22, 2011 2:04 PM
To: John Shelley
Subject: RE: Verhoeff/Masson's Stain

Your problem may be that the elastic stain should be made fresh every time, or you are differentiating in ferric chloride too long.  Does the elastic stain get mixed right before use?  Also check the expiration date of the kit. 
Shirley

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of John Shelley
Sent: Tuesday, March 22, 2011 1:59 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Verhoeff/Masson's Stain

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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