Re: hair cell IHC

<< Previous Message | Next Message >>
From:Philip Oshel <> (Karen Larison)
Content-Type:text/plain; charset="us-ascii" ; format="flowed"


I think a good part of the problem is the "freezing slowly over 
liquid nitrogen". Plunge freeze into liquid propane or ethane in LN2 
or propane-jet freeze or, if someone in your area has an instrument, 
high-pressure freeze, then freeze-substitute. After that, I'd embed 
in one of the resins good for immuno work.


>Fellow netters:
>One of the post docs here is doing IHC on frozen sections of frog 
>sacculi.  The
>staining works well, but the morphology of the hair cells is 
>horrible.  Maybe someone
>has a suggestion on how to preserve the integrity of these cells though the
>cryosectioning/staining process.  We embed the sacculus in agar, 
>sink in 30% sucrose,
>and freeze slowly over liquid nitrogen.  For most small specimens we 
>work with, this
>procedure works well, but the sacculus seems to be more 
>picky-you-nish.  So if anyone
>out there works with this organ, and has ideas on how to improve our 
>morphology, I'd
>love to hear from you.
>Karen in Oregon

Philip Oshel
Supervisor, AMFSC and BBPIC
Dept. of Animal Health and Biomedical Sciences
University of Wisconsin
1656 Linden Drive
Madison,  WI  53706-1581
voice: (608) 263-4162
fax: (608) 262-7420 (dept. fax)

<< Previous Message | Next Message >>