[Histonet] help!!

[Tony Henwood (SCHN)]

You can use DAB but the issue of endogenous peroxidase will rear its ugly head.

And I suppose IF being around since 1955, when Mellors first applied the technique to renal tissue, it has a long history of diagnostic application that is hard to replace.

There are other enzymes that could be used that, not being present in human tissue, would not require endogenous enzyme blocking for example glucose oxidase. The advantage of this enzyme is that there is no endogenous glucose oxidase activity in mammalian tissues.

Weening, J. J., & Jennette, J. C. (2012). Historical milestones in renal pathology. Virchows Archiv, 461(1), 3-11.

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children’s Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

From: Allan Wang [mailto:allanvv at gmail.com]
Sent: Tuesday, 18 April 2017 5:51 PM
To: Tony Henwood (SCHN)
Cc: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] help!!

Tim and Tony,

Why couldn't DAB be used on frozen sections in your example?

Allan

On Mon, Apr 17, 2017 at 9:03 PM, Tony Henwood (SCHN) via Histonet <histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>> wrote:
Hi Bianca,
Well for most Pathology departments, Immunofluorescence (IF) is used for Renal and Skin biopsies; looking for human Immunoglobulin (Igs) deposition on basement membranes. The advantage here is not so much the fluorescence, but that we use unfixed frozen sections. The buffer rinse before antibody application, removes un-bound serum immunoglobulins, leaving any pathological bound Igs for the IF antibody to bind to. This gives a clean result.

If one would do IF on formalin-fixed paraffin sections of renal or skin biopsies, you would find heavy background due to the fixative cross-linking serum Igs to tissue and cells (which would usually be removed by the buffer rinse if unfixed frozen sections were used - see above).

IF, apart from being a historic method, also does not suffer from endogenous peroxidase that would need to be blocked if peroxidase was used in place of fluorescence.



Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

-----Original Message-----
From: Blanca Lopez via Histonet [mailto:histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>]
Sent: Thursday, 13 April 2017 11:10 PM
To: histonet at lists.utsouthwestern.edu<mailto:histonet at lists.utsouthwestern.edu>
Subject: [Histonet] help!!
Hello!
I just need a help with a simple question...Is anyone can explain me what is the purpose between performing immunohistochemistry and Immunofluorescence?
Thanks  :)

Blanca Lopez
Histotech (ASCP)
UTSW Tissue Resource K1.210
Simmons Comprehensive Cancer Center
UT Southwestern Medical Center
Telephone: 214-648-7598<tel:214-648-7598>
Email: Blanca.Lopez at utsouthwestern.edu<mailto:Blanca.Lopez at utsouthwestern.edu>


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