Re: [Histonet] Yeh glad to be back & ??
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"
To: "'Liz Chlipala'" ; "'Amy Porter'" ;
"'Histonet'"
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@lists.utsouthwestern.edu
> [mailto:histonet-bounces@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@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@lists.utsouthwestern.edu
> [mailto:histonet-bounces@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@msu.edu
> www.humanpathology.msu.edu
>
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