[Histonet] Mouse Brain Processing Help

[Laura Tarwater]

I need some advice on processing mouse brain.  We have an outside lab using
our automatic processor and embedding equipment and has been having issues
section their mouse brain.  I was informed that the sections are extremely
brittle and rolling but mostly just the paraffin and not the tissue
itself.  They are seeing the brain tissue separate from the paraffin once
it is the water bath.

Had better results when placing a soaked gauze on the surface of the block
instead of chilling or soaking block to prevent swelling of tissue.

Here is the protocol they are using for harvesting and processing:
1.  Tissue perfused to 4% PFA at time of sacrifice
     Brain is cut in half at this point and separated
2.  Tissue placed in cassette in 10% NBF for 24hrs
3.  Then placed in 70% ETOH until able to process (usually the next day)
4.  Processing Schedule (all done with vacuum)
   30min 70%  ETOH
   30min 80%  ETOH
   30min 90%  ETOH
   30min 100% ETOH
   30min 100% ETOH
   30min 100% ETOH
   30min xylene
   30min xylen
   5min empty tank to remove excess xylene
   30min paraffin
   30min paraffin

I am not a histotech so any advice you can give me would be great.

Thanks,


*Laura Tarwater*

*Tissue Bank Technician*

*Harper Cancer Research Institute*


*University of Notre Dame574-631-2562*
*tarwater.1 at nd.edu <tarwater.1 at nd.edu>*