[Histonet] IHC on Frozen Tissue

[Sheila Fonner]

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:shive003 <@t> 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" <sfonner <@t> labpath.com>
To: "'Richard Yeo'" <ryeo <@t> wchosp.org>; <histonet <@t> 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:ryeo <@t> 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-bounces <@t> lists.utsouthwestern.edu on behalf of Sheila Fonner
> Sent: Tue 8/16/2011 7:44 AM
> To: 'Yahoo'; histonet <@t> 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.tournear <@t> 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" <sfonner <@t> labpath.com> wrote:
>
>> Thank you.
>>
>>
>> -----Original Message-----
>> From: Mierow, Brett T. [mailto:Brett.Mierow <@t> 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.mierow <@t> essentiahealth.org
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sheila
>> Fonner
>> Sent: Tuesday, August 16, 2011 5:41 AM
>> To: histonet <@t> 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
>>
>>
>>
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>
>
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