RE: [Histonet] Cresyl Violet stain for bird brains
Cresyl Violet Solution: 0.02g dye, 100ml Distilled water, 0.25ml glacial
acetic acid.
-----Original Message-----
From: Brandon Munk [mailto:bmunk@uwyo.edu]
Sent: Monday, August 08, 2005 1:02 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Cresyl Violet stain for bird brains
I am having trouble obtaining an acceptable Nissl stain of passerine brains
using cresyl violet. The birds were transcardially perfused, brains removed,
postfixed and cryoprotected before sectioning at 40 um on a sledge
microtome. The protocol I am using was originally used for rat brains and is
as follow:
ddH20 (few dips)
cresyl violet (5-10 min)
ddH20 (rinse excess stain)
70% etOH (few dips)
95% etOH (few dips)
95% etOH + acetic acid (few dips)
ddH20 (few dips)
95% etOH (few dips)
100% etOH (few dips)
repeat previous steps as necessary
Xylene (one dip)
coverslip
I am unsure as to the concentration of cresyl violet I am using and it has
sat in a fridge for >2 yrs (although I have been assured that the age should
not be a problem). I have tried playing with the protocol quite a bit,
varying time in cresyl violet or how many times I dehydrate and
differentiate. I am getting near my wits end and still cannot get a good
stain. All my sections look as if they are washed out and I often have some
stain 'smeared' under the cover slip after dipping in xylene.
Can anyone help me? Thanks in advance!
Brandon
_____
Brandon Munk
Department of Zoology and Physiology
University of Wyoming
PO Box 3166
Laramie, WY 82071
_____
bmunk@uwyo.edu
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mai
<< Previous Message | Next Message >>