Immunofluorescence (IF) or cell imaging techniques rely on the use of antibodies to label a specific target antigen with a fluorescent dye (also called fluorophores or fluorochromes) such as fluorescein isothiocyanate (FITC).
2. Introduction
Immunofluorescence (IF) or cell imaging techniques
rely on the use of antibodies to label a specific target antigen
with a fluorescent dye (also called fluorophores or
fluorochromes) such as fluorescein isothiocyanate (FITC).
Fluorochromes are dyes which have the ability to absorb
the short wavelength UV radiation and emit light of longer
wavelength fluorescence ( visible green light).
Examples : FITC, Rhodamine
3. There are two ways of doing IF staining:
Direct immunofluorescence
Indirect immunofluorescence
Direct immunofluorescence: Direct IF uses a single
antibody directed against the target of interest. The
primary antibody is directly conjugated to a fluorophore.
4. What is direct immunofluorescence?
Its applications are Clinical and Research.
Direct immunofluorescence (DIF) is a technique used in the laboratory to
diagnose diseases of the skin, kidney, and other organ systems.
DIF involves the application of antibody–fluorophore conjugate molecules to
samples of patient tissue obtained from biopsies
5.
6. Indirect
immunofluorescence:
Indirect IF uses two
antibodies. The primary
antibody is unconjugated
and a fluorophore-
conjugated secondary
antibody directed against
the primary antibody is
used for detection.
8. We want to find anti-Toxoplasma antibodies in the patients
So
What should be our method?
1. Direct immunofluorescence
2. Indirect immunofluorescence
11. TORCH Infections. Toxoplasmosis, Other (syphilis, varicella-zoster,
parvovirus B19), Rubella, Cytomegalovirus (CMV), and Herpes infections.
IgM and IgG antibodies Identified in pregnant
women.
IgM only is requested in baby …
