[Histonet] need to section -80C frozen tissue on sliding microtome

[Alan Bright]

Dear Maria,

Although I have a broad knowledge of cryostats & microtomes and some
useful sectioning techniques plus the fact I have not seen any answers
yet to your problem. Here are my two penny's worth followed after your
text with _ _ _ _

Now, my question is - if I need to use the sliding microtome to  
section these -80C frozen tissues-
how do I proceed with them so I can section them?????????? They are  
already frozen, but I
still need to attach them to the sliding tissue stage - how? _ _ _ _As
the specimen is frozen and the stage should be too, you will be unable
to use embedding medium to adhere the specimen to the tissue stage of
the microtome. Therefore you will have to use water from a syringe or
pipette and be very quick in placing the specimen in place prior to the
water freezing.

And,  they maybe too cold to section??_ _ _ _ Yes they will be too cold,
so you need to measure the temperature of the specimen with a
thermometer or electronic probe until it rises to the desired
temperature,  this is the only way you will know accurately, as it will
always be a different timing unless the specimens are all the same size,
the tissue stage is always at the same temperature and the ambient too.


Best Regards

Alan Bright

Bright Instrument Co.Ltd.
St Margaret's Way
Huntingdon
Cambridgeshire
PE29 6EU
England

Tel No:+44 (0)1480 454528
Fax No:+44 (0)1480 456031
Email: abright <@t> brightinstruments.com
Web Site: www.brightinstruments.com
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-----Original Message-----
From: Maria Mejia [mailto:mbmphoto <@t> gmail.com] 
Sent: 27 April 2006 04:00
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] need to section -80C frozen tissue on sliding
microtome


My question is a bit odd, but here goes. Over the years, I've gained  
a lot of experience
sectioning fixed brain on the sliding microtome. These specimens were  
always frozen from
a 30% sucrose/PBS solution & sectioned on the microtome.

Now, for the past 2 months I've been working in a new lab that has a  
very large tissue
bank. All of the primate tissue is fixed in zamboni fixative and they  
go through the usual
30% sucrose solution before they are snap frozen and stored in a -80  
C sub zero freezer.

If I need to cryostat section this type of frozen tissue, I know that  
once I remove the tissue
from the -80C storage area I have to leave it in the cryostat at  
least 1 hour before sectioning
it.

Now, my question is - if I need to use the sliding microtome to  
section these -80C frozen tissues-
how do I proceed with them so I can section them?????????? They are  
already frozen, but I
still need to attach them to the sliding tissue stage - how? And,  
they maybe too cold to section??

Please any suggestions, and tips you can provide will be greatly  
appreciated.

Yours

Maria Bartola Mejia
Department of Neurosurgery
University of California San Francisco
San Francisco, CA 94103

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