gluteraldehyde fixation with Re: [Histonet] Rat brains

From:Gayle Callis

Question:  is he planning to do EM on these tissues? If he had used 
paraformaldehyde, your sectioning would be much easier -  but if he is also 
doing EM - you can try some things to counteract the dry/brittle brain.

  In general, gluteraldehyde creates some friable, brittle tissues when 
these are processed and embedded in paraffin. Check to see if you are 
processing too long as long dehydrations, etc and heat from paraffin 
contributes to the problem.

A good long soak in ice water or solid ice block with water on top after 
trimming might help but be sure to not retrim the soak away, you have to 
get the first sections off the knife.  A warm water or room temperature 
soak the going onto ice water for a time may help too, brain sometimes 
needs a bit of help.

At 01:01 PM 7/23/2005, you wrote:
>Please help...
>A research project for our neuropathologist includes rat brains.  These 
>are extremely difficult to section as he profuses the whole rat with 
>glutaraldehyde then takes brain samples.  I have tried adding alcohol to 
>my water bath but these sections still have many, many wrinkles.  Can 
>anyone offer suggestions?  I would be so grateful for any and all advice.

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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