[Histonet] to cut or not to cut mouse brains

2011-11-17 Thread McLaughlin, Terry 
Hello All,
I am having trouble cutting frozen mouse brains and was wondering if
someone can offer some help. The mouse was perfused in 4% PFA , the
brain removed and kept in PFA for 22 hrs. We placed it in 30% sucrose
for 20 hrs, until the brain sank in this solution. It was frozen back by
using a culture plate floating in liquid nitrogen with OCT . 
The problem I am having is many folds, and air bubbles under the tissue.
What I have done so far:
Cut at temp of -25, then tried - 22 and -17.
Cut at 10um, then tried 16-25um.
Tried making the knife colder by adding dry ice to it for a few seconds.
Added dry ice to sample for a couple seconds.
The sample cuts fine, it appears to happen when picking up on a slide. I
tried picking up by starting at one side or end and letting tissue
spread onto slide. Did not work.
Also tried gently laying slide on top of sample and removing
gently-still folds and air bubbles. Any help would be greatly
appreciated as we have 6 more to do.
 
Signed,
Help!
Terry Kokas
 
 
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[Histonet] Help finding metal embedding molds

2011-10-25 Thread McLaughlin, Terry 
Hello All,
This is my first time posting a request. We use metal embedding molds,
and would like to order one particular size. The outer measurements are
2.5mm x 2.5mm x 
0.6 mm. We are wondering if anyone has approximately 40-50 of them we
could purchase-new or used. The reason we need this size is because most
of our research tissue fits perfectly into this size and depth. We have
18 of them and quite often need 30-60 for a particular project. We found
them from one of the vendors, but the price is quite expensive. We are
hoping someone out there can help us.
 
Much Thanks,
Terry McLaughlin 
Histology Specialist 
AI DuPont Hospital for Children 
1600 Rockland Rd. 
Wilmington, DE. 19803 
phone 302-651-6771 
fax-302-651-5010 

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