[HISTONET] fixation and storage of tissue for TEM

[Geoff McAuliffe]

Fix in buffered glutaraldehyde for 2 hours, overnight is acceptable. 
After multiple rinses, store in buffer in the refrigerator. Change the 
buffer once a week, there are things that will grow in cacodylate 
buffer. You may complete osmication if you want, then rinse and store in 
buffer. I would not store the tissue indefinitely in anything. Do NOT 
store the tissue in 70% ethanol, either before or after osmium, ethanol 
has been shown to extract cytoplasm from fixed tissues.
The texts John Kiernan recommends are excellent.

Geoff

Nicholas David Evans wrote:
> Dear all,
>
>  
>
> I would like to fix and preserve mouse skin tissue for processing for
> transmission electron microscopy (TEM) at a later date. I was wondering
> whether I can store tissue following fixation overnight in 2.5%
> gluteraldehyde in 0.1M sodium cacodylate buffer and, if so, whether it can
> be stored indefinitely in this buffer (or whether I need to treat with
> OsO4 and store at 70% ethanol)?
>
>  
>
> Thanks and best wishes
>
> Nick
>
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>
>   


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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583 
mcauliff <@t> umdnj.edu
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