[Histonet] brain histology

[I-sanna Gibbons]

I actually used 10% formalin initially.  First I perfused the bat transcardially, then removed the brain and placed it in the same fixative. That was the first batch.  For the second batch of bats, I removed the brain (without prior perfusion) immediately after the animal had expired and placed it in Carnoy's.
  Dr. Cross, glad to meet you as well.  I was actually thinking of pursuing pathology after spending 3 years in the anatomy department post-graduation.  Are there any vet pathology residencies you would recommend?
  I-sanna

Cheryl Cross <ccross6032 <@t> aol.com> wrote:
        

  Hi Dr. Gibbons - 
  

  Great to see a fellow vet posting!! I do some neurotox stuff in mice. What makes for a really nice FFPE section is, if you can, perfuse fixing the rodents, then removing the calvarium overlying the brain, and letting the brain sit in the skull in your fixative of choice (I use 10% NBF). Letting the brain sit for 24 hours in the fix allows you to avoid those dreaded dark angular neurons. After sitting for 24 hours I take out the brain and section routinely ... if I can't section immediately and immunohistochemistry will be performed i switch it to 70% EtOH ... but sitting in the alcohol makes the tissue very very firm. If immunohistochemistry isn't an issue I allow the brain or organs to remain in the 10% NBF. 
  

  Hope this helps, let me know if you need a protocol for perfusion, etc. 
  

  Cheryl
  

      Cheryl Cross, DVM, Dipl. ACVP
  Researcher
  University Corporation for Atmospheric Research
  College of Veterinary Medicine
  University of Tennessee Department of Pathology
  2407 River Drive, Room A201
  Knoxville, TN 37996-4542
  (423) 967-2724
  fax: 865-974-5616
  ccross <@t> ucar.edu



  




    On May 9, 2007, at 4:50 PM, Rene J Buesa wrote:

    I think is too much time in alcohol(s). Storing in absolute EthOL is not a good practice.
    Try to reduce this aspect and check for results.
    René J.
  

  I-sanna Gibbons <trinimaican2501 <@t> yahoo.com> wrote:
    I am having a bit of trouble obtaining good quality slides of bat brains. The brains are fixed in Carnoy's, stored in absolute alcohol, processed routinely, infiltrated with paraffin, sectioned at 8 microns and stained using cresyl fast violet. The aim is to record the cytoarchitecture. The resulting slides show poor tissue integrity with extensive cracking. Can anyone suggest how this can be avoided? I was thinking maybe a double-embedding method for whole brains (10-15 mm) and thicker sections (25-30 microns)?
  

  Thanking you in advance
  

  I-sanna Gibbons, DVM
  Veterinary Anatomy
  School of Veterinary Medicine
  The University of the West Indies
  Trinidad and Tobago, W.I.
  

  

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