[Histonet] help!!

[Tony Henwood (SCHN)]

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] 
Sent: Thursday, 13 April 2017 11:10 PM
To: 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
Email: Blanca.Lopez at utsouthwestern.edu


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