Hi Histonetters-

I am working on a research project which involves Nissl staining (with 
cresyl violet) mouse brains in order to examine the hippocampus.  It is 
kind of a side project (my lab is actually a neurophysiology lab so 
histology equipment is very limited), so no one really has much experience, 
and I am pretty much self-taught.  It is not going well at all and I would 
love some help. Here's basically what I've been doing (I expect most of it 
is incorrect!)

I take out the brain and fix it in 4% paraformaldehyde for 24 hours.
Then I've been slicing each hemisphere on a manual tissue slicer at 
approximately 50-70 microns
I mount the slices on chrom-alum-gelatin subbed slides and let dry at least 
overnight
Then I stain using cresyl violet and dehydrate in graded alcohols (50, 70, 
85, 95, 100) and coverslip with Permount.

My main problem is that all of my slices crack and virtually disintegrate 
when I am coverslipping.  I think this might be because they are too thick, 
but I am not sure.

Basically, I just need to be able to quickly compare the structure of the 
hippocampus in different mouse subtypes.  If you have any tips or think you 
can solve my slice cracking problem, please let me know!

Thanks,
Michelle Sotak
Research Assistant
Trommer Laboratory
Evanston-Northwestern Research Institute- Neurology
m-sotak@northwestern.edu

Michelle Sotak
Research Assistant
Trommer Laboratory
Evanston-Northwestern Research Institute- Neurology
2650 Ridge Avenue
Evanston, IL 60201
(847) 570-2289   fax (847) 733-5256
m-sotak@northwestern.edu