[Histonet] Immunoperoxidase Protocol on Cytospin

[Cartun, Richard]

Since you only have 1 slide I would recommend following your regular protocol that you use for formalin-fixed, paraffin-embedded tissue.  I would use heat retrieval if that's what you use for FFPE tissue.  However, if you use enzyme digestion on FFPE tissue I would "not" digest your cytospin slide.  If you don't get any immunoreactivity it could be a "False Negative" due to pre-analytical variables.  I think it can be interesting to try experiments like this, but it's not the way to do IHC testing.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology & Morphologic Proteomics Laboratory
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax




-----Original Message-----
From: Margaryan, Naira [mailto:NMargaryan at luriechildrens.org] 
Sent: Tuesday, March 14, 2017 3:24 PM
To: Cartun, Richard
Subject: RE: Immunoperoxidase Protocol on Cytospin

cytoplasmic staining for Nodal protein.

thanks
________________________________________
From: Cartun, Richard [Richard.Cartun at hhchealth.org]
Sent: Tuesday, March 14, 2017 2:13 PM
To: Margaryan, Naira; histonet-bounces at lists.utsouthwestern.edu
Subject: RE: Immunoperoxidase Protocol on Cytospin

What is the target?

Richard

Richard W. Cartun, MS, PhD
Director, Histology & The Martin M. Berman, MD Immunopathology & Morphologic Proteomics Laboratory Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596
(860) 545-2204 Fax




-----Original Message-----
From: Margaryan, Naira via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, March 14, 2017 2:51 PM
To: histonet-bounces at lists.utsouthwestern.edu
Cc: histonet-request
Subject: [Histonet] Immunoperoxidase Protocol on Cytospin
Importance: High

Dear histonetters:

I never done ICC.

A scientist brought me a slide with Cytospined cells on for IHC.

I am going to fix with 10%NBF (it is only what I have now) then wash with TBST.

May I skip AR step?
May I use my usual IHC protocol and reagents which I usually use for FFPE tissue: blocks with H2O2, avidin/biotin/PB with TBST wash buffer????

Thanks in advance,
Naira


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