[Histonet] brazilin

[Paula Pierce]

Hi,
 
when I do Lac Z, I counterstain with nuclear fast red, rinse in water, then use one of the aqueous mounting medias that have replaced Crystal Mount. 
http://www.emsdiasum.com/microscopy/technical/datasheet/17985-12.aspx
 
Place enough to cover the section, drain off the excess, completely air dry, and then you can permanently coverslip with resin. This is also the procedure I use for AEC and even Oil Red O.
Crystal Mount forms a thin "plastic" protective film over the section.

Paula K. Pierce, HTL(ASCP)HT
President
Excalibur Pathology, Inc.
8901 S. Santa Fe, Suite G
Oklahoma City, OK 73139
405-759-3953 Lab
405-759-7513 Fax
www.excaliburpathology.com

From: Geoff McAuliffe <mcauliff <@t> umdnj.edu>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Tuesday, August 30, 2011 11:34 AM
Subject: Re: [Histonet] brazilin

Hi Vanessa:

I tried "Brizilliant" several years ago. I purchased the dry powder 
since I have been mixing my own stains since the 1970's. I even called 
the vendor as I was unsure about the identity of some of the 
ingredients. The results on formalin fixed, paraffin embedded rodent 
tissues were disappointing, a dull orange not bright red. You might have 
better results with the pre-mixed stain. Perhaps they wills send you a 
small sample to try on your material.

Geoff

On 8/30/2011 9:15 AM, Vanessa Orsini wrote:
> Hi everyone,
>
> I'm Vanessa and i'm quite new in the histology field so i need some help...
>
> i'm having problems with a LacZ staining on mouse fresh frozen tissue. I managed to get a nice staining also if I think it's a bit variable but I cannot find a good counterstain!!
> I tried the nuclear fast red but it's not stable in an aqua-based mounting medium and if I do the acohol row before mounting with pertex the morphology of the sections is damaged and the lacz staining is a bit washed away (a problem if the Bgal expression is low)
> I then tried with ematoxilin, the staining is stable but it's more difficult to discriminate between the blue of the lacZ and the blue of ematoxilin.
> I find out that the brazilin is a red ematoxilin so i decided to try this one but once the powder arrived i didn't received any instruction for dilution with it I couldn't find anything on the net.
>
> Does someone have experience with brazilin?
> or do you know some other red counterstaining that I could use?
>
>
> Thanks a lot
>
> Vanessa
>
>
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-- 
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583
mcauliff <@t> umdnj.edu
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