Jan, Thanks, I really appreciate the help. This is my first attempt, so we'll see how it turns out. :)
Sheila -----Original Message----- From: Jan Shivers [mailto:shive...@umn.edu] Sent: Tuesday, August 16, 2011 9:21 AM To: Sheila Fonner Subject: Re: [Histonet] IHC on Frozen Tissue Sheila, On frozens, there should be no need to do tissue conditioning/pretreatment, since there are no fixation-caused aldehyde bonds to break. Bring your frozens to room temperature slowly, immerse in your rinse buffer for a few minutes, then proceed with your IHC staining protocol. With frozens, since the tissue can at times be more fragile, I always do the H2O2 step AFTER the primary antibody (but before the HRP-conjugated linking Ab; you don't want to quench your substrate enzyme) and I use 0.3% H2O2 instead of 3%. Jan Shivers IHC Section Head UMN Vet Diag Lab ----- Original Message ----- From: "Sheila Fonner" <sfon...@labpath.com> To: "'Richard Yeo'" <r...@wchosp.org>; <histonet@lists.utsouthwestern.edu> Sent: Tuesday, August 16, 2011 7:24 AM Subject: RE: [Histonet] IHC on Frozen Tissue > Rich, > > > > Isn't the cell conditioning (retrieval) mostly done for tissues that have > been in formalin? If the tissue is fresh (frozen), it has not seen > formalin. That is the reason I questioned the cell conditioning step. > > > > Thank you for your help. > > Sheila > > > > > > > > From: Richard Yeo [mailto:r...@wchosp.org] > Sent: Tuesday, August 16, 2011 7:54 AM > To: Sheila Fonner > Subject: RE: [Histonet] IHC on Frozen Tissue > > > > Sheila, > > > > The cell conditioning step is for retrieval. It will depend on what > antibody > you are going to run. If you use it for Depar. you will also use it > wiith-out. > > > > > > Rich Y > > > > _____ > > From: histonet-boun...@lists.utsouthwestern.edu on behalf of Sheila Fonner > Sent: Tue 8/16/2011 7:44 AM > To: 'Yahoo'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] IHC on Frozen Tissue > > Does anyone else leave out the cell conditioning step? > > > -----Original Message----- > From: Yahoo [mailto:kim.tourn...@yahoo.com] > Sent: Tuesday, August 16, 2011 7:41 AM > To: Sheila Fonner > Subject: Re: [Histonet] IHC on Frozen Tissue > > I would leave out the pretreatment step as well i.e. Enzyme, CC1/CC2. > > Sent from the iPhone of Kim Tournear > > On Aug 16, 2011, at 6:03 AM, "Sheila Fonner" <sfon...@labpath.com> wrote: > >> Thank you. >> >> >> -----Original Message----- >> From: Mierow, Brett T. [mailto:brett.mie...@essentiahealth.org] >> Sent: Tuesday, August 16, 2011 7:02 AM >> To: Sheila Fonner >> Subject: RE: [Histonet] IHC on Frozen Tissue >> >> You are correct, leave out the depar. step. >> >> >> Brett Mierow, HT (ASCP) >> Histology specialist >> Essentia Health >> SMDC Laboratory >> 407 East 3rd Street, Duluth, MN 54880 >> (218) 786-4510 | brett.mie...@essentiahealth.org >> >> -----Original Message----- >> From: histonet-boun...@lists.utsouthwestern.edu >> [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sheila >> Fonner >> Sent: Tuesday, August 16, 2011 5:41 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] IHC on Frozen Tissue >> >> Good morning everyone, >> >> >> >> I was wondering if anyone could help me with a protocol change for the >> Ventana Ultra. I have a pathologist that is requesting an HSVI, HSVll, >> and Zoster Virus on frozen tissue that was received for IF studies. I >> know it's not Monday, but it is early, and I was hoping for some >> information about how to change the protocol for frozen vs. paraffin >> tissue. Do I just need to delete the depar. step and leave everything >> else the same? I appreciate any help you could give. >> >> >> >> Thanks, >> >> Sheila >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
