[Histonet] Yeh glad to be back & ??

[Amy Porter]

Thanks Patsy - Are you using Caspase 3 ?  or another Caspase for that  and 
is it active or cleaved - just curious.  I think I am going to have trouble 
all the way around.  We are trying to do immuno on mouse tibias that have 
been acid decalcified.  After several postings I am trying to look at 
alternative decalcification methods, I have been able to obtain some 
forelimbs from mice just to work with the decaling and processing.  Any 
pointers that you would have would be greatly appreciated.  Amy
----- Original Message ----- 
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
To: "'Liz Chlipala'" <liz <@t> premierlab.com>; "'Amy Porter'" <portera <@t> msu.edu>; 
"'Histonet'" <histonet <@t> pathology.swmed.edu>
Sent: Friday, September 22, 2006 3:43 PM
Subject: RE: [Histonet] Yeh glad to be back & ??


> Amy,
> I agree with Liz on this, it is hard enough with Tunnel to distinguish
> between necrosis and apoptosis without adding decal.  I much prefer 
> caspase,
> you can count on the positive cells being just those under going 
> apoptosis.
> Patsy
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Liz 
> Chlipala
> Sent: Thursday, September 21, 2006 9:45 AM
> To: 'Amy Porter'; 'Histonet'
> Subject: RE: [Histonet] Yeh glad to be back & ??
>
> Amy
>
> I did this back a few years ago and found that the process of formic acid
> decalcification caused all the nuclei to be positive.  From what I 
> remember
> the use of formic acid is not compatible with the tunel technique.  Why
> don't you try using cleaved caspase 3 immunohistochemistry instead.
>
> Liz
>
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Manager
> Premier Laboratory, LLC
> P.O. Box 18592
> Boulder, CO 80308
> phone (303) 735-5001
> fax (303) 735-3540
> liz <@t> premierlab.com
> www.premierlab.com
>
> Ship to Address:
>
> Premier Laboratory, LLC
> University of Colorado at Boulder
> MCDB, Room A3B40
> Boulder, CO 80309
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amy Porter
> Sent: Thursday, September 21, 2006 7:09 AM
> To: Histonet
> Subject: [Histonet] Yeh glad to be back & ??
>
> Is anyone out there doing Tunel on FFPE acid decalcifed sections.  I am
> trying to stain for apoptotic cells using a Tunel kit on decalcified mouse
> femurs.  Not having much success, if anyone has any tips or do's / don'ts 
> I
> would really appreciate the information.  Thanks in advance,
>
>
> Amy S. Porter, HT(ASCP) QIHC - Supervisor
> Michigan State University
> Investigative HistoPathology Laboratory
> Department of Physiology / Division of Human Pathology
> 2100 Biomedical Physical Sciences Bldg. Room #2133
> East Lansing, MI  48824-3320
> Phone:  (517) 355-6475 ext 1480 / Fax:  (517) 432-1368
> Email:  portera <@t> msu.edu
> www.humanpathology.msu.edu
>
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