Re: Frozen tissues

From:Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of histonet)

Whoops, I said that and it is incorrect, for sectioning liver, I RAISE the
temperature to -16C (lower number means higher temperature in cryostat!)
Must be holiday time!

At 08:02 AM 8/30/00 +0000, you wrote:
>I never cryopreserve rodent tissues before snap freezing unless they are
>prefixed in PFA or NBF before freezing.  Just lower the temp on cryostat to
>-16C and go for it.  You can try different fixatives, PFA, NBF,  acetone 10
>min on air dried sections 30 min to overnight), RT alcohol 5 min fix on air
>dried sections or acetone/alcohol RT fix 5 min on air dried.
>
>Do not defrost the tissue, cut the frozens on blocks as they are, put
>tissue on block with OCT, can even surround tissue with OCT then let it
>freeze.
>
>
>
>08:11 AM 8/31/00 -0600, you wrote:
>>Greetings everyone!
>>
>>I just got a call from the guy I do HBV IHC for and he has a project for
>>me.  Th problem is that all his tissues (transgenic mouse livers) are
>>frozen, the tech who froze them put them in lq N2 but DID NOT use a
>>cryopreservant.  We want to do frozen sections on these and perform HBV
>>IHC on them.  Is it best to defrost them and fix in BNF or leave them
>>frozen?  The concern here is all the other work I've done has been on
>>BNF fixed & paraffin embedded tissues and we would like some modicum of
>>consistency.
>>
>>thanks
>>
>>Connie McManus
>>
>>
>>
>>
>>
>Gayle Callis
>Veterinary Molecular Biology
>Montana State University
>Bozeman MT 59717-3610
>406 994-4705
>406 994-4303
>
>
>
Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303




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