[Histonet] 20 micron sections Cresyl Violet Acetate stained

From:"Patrick Laurie"

Hi Histonet,

My group is interested in doing an anterior horn alpha motor neuron
count of ALS pts vs control.  There is considerable literature about
these techniques, which has been most helpful, but the devil is in the

Most papers use 20um sections of the ventral horn stained with cresyl
violet acetate, or iron-chromoxane cyanine R method for myelin
(counterstained with .5% neutral red) (Dr. Kiernan's 1991 paper), or one
of many other methods for staining.  I have 3 questions.  First of all,
a technical question, how are 20um sections kept on the slides during
staining?  I have done some experimenting, and found it very difficult
to keep them on.  

Secondly, how do the thick sections stain?  The cresyl violet using will
not differentiate very well (using the regressive version), and the H&E
is too dark, I have yet to use any other method. 

And lastly, why 20um?  Conventional neural histology seems to be done
thicker, but usually not that thick.  What would be the benefit for
having sections that thick?  

Thanks Histonet,

Patrick Laurie, HT (ASCP)
Neurogenomics Laboratory
Benaroya Research Institute
1201 9th Ave
Seattle, Wa  98101

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