[Histonet] Cresyl Violet Counterstain of LFB

Amos Brooks amosbrooks <@t> gmail.com
Thu Jun 4 18:19:02 CDT 2009

Hi Nicole,
   Why bother dehydrating in ETOH then? Just rinse then air dry the
sections. I had the same problem recently and this is what I do now to avoid
the whole destaining problem. Once the slides are air dried, you can put
them directly in xylene.

Message: 12
Date: Thu, 4 Jun 2009 09:54:58 -0400
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>
       <E4D68832BCDC974CA4302A47FD8B0C80014D4FAC48 <@t> NIHMLBX06.nih.gov>
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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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