FW: Cresyl Violet Staining

From:Lorraine Masse <lmasse@biosupport.com>

> I am using Cresyl Violet Acetate to stain serial sections of macaque
> brain.  This tissue has been fixed in formalin for over 6 months, frozen
> sectioned at 40 microns, and placed in PBS with 0.05% Sodium Azide before
> mounting on slides.  I have been running into problems with fat
> displacement when I mount the slides onto coverslips.  The fat tends to
> spread all over the slide and causes problems when viewing the slides
> under the microscope.  The protocol for staining calls for 2 minutes of
> hydration in deionized water, 1.5 minutes in the Cresyl Violet Stain, 4
> rinses in tap water for 30 seconds each, run through alcohol at 80, 95,
> 100, and 100 percent for 30 seconds each, 2 minutes in xylene twice, and
> coverslip.  I have tried hydrating the tissue in alcohol at 100, 95, 70,
> 50, and 35 percent before starting the deionized water step,  which caused
> the stain to be too dark for counting neurons.  I have also tried fixing
> the tissue in 40% formalin before starting the deionized water step, which
> discolors the stain.  Do you have any other ideas that might be
> successful?
> Thank you,
> Sarah McGrath

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