[Histonet] help!!

[Morken, Timothy]

Blanca, immunofluorescence (IF) is a subset of immunochemistry. Immunohistochemistry is also a subset of immunochemistry. There is some overlap between the two.

Immunohistochemistry denotes  immunochemistry done on tissue sections ("-histo-" =" tissue"). But we can also use other enzymes to label the antibodies for immunohistochemistry (peroxidase, alkaline phosphatase, etc).


IF is just one of many methods of labeling the antibodies with a visual label. Others are peroxidase and alkaline phosphatase.


Generally IF is done on "fresh" cells or tissue. For tissue it is normally frozen tissue. 

IF can be done on cells (ie, immunocytochemistry) either on slides (smears, various preparations) or in solution as with flow cytometry - the cells are labeled with fluorescent-labeled antibodies and sorted by color (or no color). 

Generally the IF method is faster to perform because there is no processing beyond freezing the tissue. In the past IF was also more sensitive due to dark field microscopy in the fluorescence microscope. With the advent of various methods to amplify the signal (avidin -biotin, polymers with multiple enzymes) the peroxidase methods are just as sensitive, if not more so.

But fresh or frozen tissue has the advantage of the epitopes remaining unfixed, especially by formalin - which can mask the antigen from the antibody. Some antibodies do not work well on formalin-fixed tissue, even if antigen retrieval is used, so frozen tissue or cells are the best option. 



Tim Morken
Pathology Site Manager, Parnassus 
Supervisor, Electron Microscopy/Neuromuscular Special Studies
Department of Pathology
UC San Francisco Medical Center





-----Original Message-----
From: Blanca Lopez via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Thursday, April 13, 2017 6:10 AM
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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