[HISTONET] fixation and storage of tissue for TEM

[Nicholas David Evans]

Thanks for the helpful replies and for the reading suggestions. We are
fixing for a collaborator who'll do the embedding, so I just wanted to
check on the best way to store/transport. This makes it clearer.

Best wishes
Nick

-----Original Message-----
From: Geoff McAuliffe [mailto:mcauliff <@t> umdnj.edu] 
Sent: Wednesday, December 09, 2009 11:16 AM
To: Nicholas David Evans
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [HISTONET] fixation and storage of tissue for TEM

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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Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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