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@lists.utsouthwestern.edu]
Sent: Thursday, April 13, 2017 6:10 AM
To: histonet@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.lo...@utsouthwestern.edu
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