RE: [Histonet] IHC on Frozen Tissue
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" To: "'Richard Yeo'" ; 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" 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
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" 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
RE: [Histonet] IHC on Frozen Tissue
I leave out the cell conditioning step as well. Sandra Sandra Esparza HT(ASCP)QIHC Lead Technologist Dell Children's Medical Center of Central Texas 512-324- x87061 sespa...@seton.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 6:44 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" 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
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" 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
RE: [Histonet] IHC on Frozen Tissue
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
