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immunoenhancement 365 immunofluorescence
medium containing electrolyte to identify separated antigenic
substances. This allows determination of the presence or
absence of immunoglobulin molecules of various classes in
a serum sample. One percent agar containing electrolyte is
layered onto microscope slides and allowed to gel, and patterns
of appropriate troughs and wells are cut in the solidified
medium. Antigen to be identified is placed in the circular
wells cut into the agar medium. This is followed by electrophoresis
that permits separation of the antigenic components
according to their electrophoretic mobility. Antiserum is
placed in a long trough in the center of the slide. After antibody
has diffused through the agar toward each separated
antigen, precipitin arcs form where the antigen and antibody
interact. Abnormal amounts of immunoglobulins result in
changes in the shape and position of precipitin arcs when
compared with the arcs formed by antibody against normal
human serum components. With monoclonal gammopathies,
the arcs become broad, bulged, and displaced. The absence
of immunoglobulin classes such as those found in certain
immunodeficiencies can also be detected with IEP.
immunoenhancement
Increasing or contributing to the level of immune response by
various specific and nonspecific means such as immunization.
Immunoferritin
Immunoferritin method. A ferritin-conjugated anti-A globulin treated
group A red cell. The binding of ferritin particles to the surface of the
erythrocyte denotes the site of the antigen.
immunoferritin method
A technique to aid detection by electron microscopy of
sites where antibody interacts with antigen of cells and tissues.
Immunoglobulin may be conjugated with ferritin, an
electron-dense marker, without altering its immunological
reactivity. These ferritin-labeled antibodies localize molecules
of antigen in subcellular areas. Electron-dense ferritin
permits visualization of antibody binding to homologous
antigen in cells and tissues by electron microscopy. In addition
to ferritin, horseradish-peroxidase-labeled antibodies
may also be adapted for use in immunoelectron microscopy.
immunoferritin technique
A method that involves the labeling of antibody molecules
with an electron-dense material (e.g., ferritin), rendering
their site of attachment to cell or tissue antigen visible when
observed by electron microscopy. In addition to this direct
method, the use of ferritin-labeled antiimmunoglobulin
can detect unlabeled antibody bound to tissue antigen by
indirect “sandwich” immunoferritin methodology.
immunofixation
A method to identify antigen in a protein mixture that has
been electrophoresed in agarose gel. Antibodies are applied
to the gel, which is then rinsed to remove excess reactants,
and the preparation is stained to reveal precipitates of antigen
and antibody.
immunofixation electrophoresis (IFE)
The application of specific antibodies to a gel in which the
antigen has been electrophoresed. Precipitation occurs at
the location where antibodies interact with the electrophoresed
homologous antigen. The method is used to identify
specific antigenic components that might otherwise be camouflaged
because of close similarity of molecules not otherwise
discernible by conventional immunoelectrophoresis.
Fluorescence antibody method showing immune deposits in
glomerulonephritis.
immunofluorescence
A method for the detection of antigen or antibody in cells
or tissue sections through the use of fluorescent labels (fluorochromes)
by fluorescent light microscopic examination.
The most commonly used fluorochromes are fluorescein
isothiocyanate, which imparts an apple-green fluorescence,
and rhodamine B isothiocyanate, which imparts a reddishorange
tint. This method, developed by Albert Coons in the
1940s, has a wide application in diagnostic medicine and
research. In addition to antigens and antibodies, complements
and other immune mediators may also be detected
by this method. It is based on the principle that, following
adsorption of light by molecules, cells or tissues dispose of
their increased energy by various means, such as emission
of light of longer wavelength. Fluorescence is the process
whereby emission is of relatively short duration (10 –6 to 10 –9
seconds) for return of the excited molecules to the ground
state. The active groups in protein that allow them to attach
fluorochromes include free amino and carboxyl groups
at the ends of each polypeptide chain, many free amino
groups and lysine side chains, many free carboxyl groups in
asparatic and glutamic acid residues, the guanidino group
of arginine, the phenolic group of tyrosine, and the amino
groups of histidine and tryptophan. Labeling antibody
I