[Histonet] Storage after perfusion of brain tissue, what is the best method?

[Charles Scouten]

They are fixed after overnight exposure.  30% sucrose is the last step.  They can be stored indefinitely.  They may be somewhat overfixed for some antibody studies, fine for anything else.  Store in sucrose on a shelf.  Yes, after sectioning, dry the slides, at least a few hours, overnight is ok.  Store for some time that way, but stain and coverslip for permanent storage.  

Be sure the tissue is frozen as fast as possible, immersion in isopentane at -80C is best, on a cold pedestal and covered with powdered dry ice is second.  Freezing in the crystat on a pedastal is unacceptable, will get swiss cheese freezing artifact damage in the tissue.


Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten <@t> myneurolab.com 
http://www.myneurolab.com 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kelly D Mcqueeney
Sent: Thursday, March 31, 2005 9:11 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Storage after perfusion of brain tissue,what is the best method?

Dear Histonetters,
I have perfused rat brain with 4% paraformaldehyde, placed in fix O/N and then added a 10% and 30% sucrose solution. What is the next step toward long-term storage of this tissue if I plan on using a cryostat to section? How would you recommend storing this tissue and what is the next step after sucrose. I will not be sectioning for a few weeks. Also, how do I store the slides after sectioning? Do I let them dry for a few hours like I do with fresh tissue?

Thanks for your help,
Kelly McQueeney

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