Re: [Histonet] museum specimens
I don't know if 'refixing' in PFA will do much to tissue that has
already seen formalin and 70% ethanol for extended periods of time. The
'best' way to proceed probably depends on how thick you want the
sections to be for the microscopic method you want to use. Paraffin
embedding will allow for the thinest sections, less than 20 microns.
Frozen/vibratome for thicker sections (20-50 microns), perhaps
nitrocellulose for really thick (100 micron) sections for 3D
reconstruction of Purkinje cells?
I would try one method on one or two brains to see how it works
before committing all of the tissue to one fate.
Andrew Iwaniuk wrote:
>I have a question regarding the preparation of museum specimens for histology.
> I recently obtained access to several museum specimens (formalin fixed and
>then preserved in 70% ethanol) that we are using for a comparative
>neuroanatomical study. We are hoping to successfully extract the brains and
>section the tissue to examine cerebellar structure. I was thinking of taking
>the brains through a graded series of ethanols to PBS, post-fixing in PFA and
>gelatin embedding them for frozen sections. Alternatives would be to leave it
>in 70% ethanol and section it on a vibratome or paraffin embed them.
>Does anyone have any experience in performing histology on museum specimens
>like these or suggestions/advice on the matter?
>Andrew N. Iwaniuk
>Post-doctoral Research Fellow
>Songbird Neuroethology Lab
>Department of Psychology
>University of Alberta
>P217 Biological Sciences Building
>Edmonton, Alberta, T6G 2E9
>Ph: +1 780 492 0323
>Fax: +1 780 492 1768
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