[Histonet] Cresyl Violet Counterstain of LFB

[Rene J Buesa]

There is no problem with that. The whole idea of washing with ethanol is to differentiate and the section will be differentiated when no more color washes out.
I don't know what artifacts you have but after sectioning there is nothing you can do. You can prevent the artifacts while sectioning and usually they are due to the time it takes to freeze the tissue and the freezing temp. also.
René J.

--- On Thu, 6/4/09, Patten, Nicole (NIH/NIAAA) [F] <pattennj <@t> mail.nih.gov> wrote:


From: Patten, Nicole (NIH/NIAAA) [F] <pattennj <@t> mail.nih.gov>
Subject: [Histonet] Cresyl Violet Counterstain of LFB
To: "'histonet <@t> lists.utsouthwestern.edu'" <histonet <@t> lists.utsouthwestern.edu>
Date: Thursday, June 4, 2009, 9:54 AM


Hi Histonet... Quick question!!

I'm doing Luxol Fast Blue staining on frozen human brain sections (1% LFB in 95% EtOH with Hydroquinone/Sodium Sulfite Differentiator) and trying to counterstain with Cresyl Violet. Following Cresyl Violet incubation (0.1% for 5') my protocol calls for me to dehydrate the tissue in ethanol (95%x2, 100%x2) but this pulls out basically all of my CV stain! Does anyone have a solution for this? 

Also, I have horrible frozen artifacts... is there anything I can do even though the tissue has already been sectioned? What about during the sectioning process?

Thanks in advance!

Nicole J. Patten
Post-Baccalaureate Fellow/IRTA
NIAAA/National Institutes of Health

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