Re: [Histonet] Best Way to obtain specific area of mouse foetal and neonatal Brain for EM and light microscopy

From:Jo Dee Fish

Dear Rowani,
Here is what I do to select specific areas of the brain for EM. 
First, cut 100um vibratome sections after the initial glutaraldehyde 
fixation.  Then process these sections through dehydration and 
infiltration with Eponate 12.  Embed them between Aclar sheets (I buy 
mine from Ted Pella) with just a small drop of resin for each 
section.  I have embedded at least 50 sections between two 8x6" 
sheets.  After they have polymerized you can look at the sections 
with an ordinary upright microscope.  You can then select specific 
areas and mount them onto empty or "blank" resin blocks (that you 
have made ahead of time).  Just remove the area you have chosen with 
a razor blade or scalpel.  You might practice first before going for 
the important and rare specimen!  I use regular gel type Super Glue 
to glue the areas to the blocks.  After they have dried you can trim 
and section just like any other EM block.
The nice benefit of this method is that the embedded sheets can be 
stored in an ordinary binder until you need them again.
This works great for me, hope it works great for you.
Take care and good luck,
Jo Dee

At 9:08 AM +0930 4/27/04, Rowani wrote:
>Yes, its electron microscopy but what I meant is to cut specific area of the
>brain (ie. hippocampus, motor cortex etc) which could then be used for  EM
>and various other purposes.
>Thank You
>Histonet mailing list


Jo Dee Fish
Research Technologist III
Gladstone Institute of Cardiovascular Disease

Telephone: (415) 695-3720
Fax: (415) 285-5632

Mailing address:
Gladstone Institutes
P.O. Box 419100
San Francisco, CA 94141-9100

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