Immunofluorescence

Immunofluorescence - Direct immunofluorescence 2. Indirect

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Immunofluorescence Sayeed Ismail Khatib Lecturer Ibn Sina National College for Medical Studies
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Introduction: Immunofluorescence is the labeling of antibodies or antigens with fluorescent dyes. This technique is sometimes used to make viral plaques more readily visible to the human eye. Immunofluorescent labeled tissue sections are studied using a fluorescence microscope. Fluorescein is a dye which emits greenish fluorescence under UV light. It can be tagged to immunoglobulin molecules.
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There are two ways of doing IF staining Direct immunofluorescence Indirect immunofluorescence 1. Direct immunofluorescence Ag is fixed on the slide Fluorescein labeled Ab’s are layered over it Slide is washed to remove unattached Ab’s Examined under UV light in an fluorescent microscope The site where the Ab attaches to its specific Ag will show apple green fluorescence Use: Direct detection of Pathogens or their Ag’s in tissues or in pathological samples
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Unformatted text preview: Direct immunofluorescence 2. Indirect immunofluorescence: Indirect test is a double-layer technique The unlabelled antibody is applied directly to the tissue substrate Treated with a fluorochrome-conjugated anti-immunoglobulin serum Advantage over direct IF Because several fluorescent anti-immunoglobulins can bind to each antibody present in the first layer, the fluorescence is brighter than the direct test. It is also more time-efficient since it is only one signal labelled reagent, the anti-immunoglobulin, is prepared during the lengthy conjugation process Indirect immunofluorescence of iron-regulated cell wall mannoprotein FIT1 of S. cerevisiae Confocal image to detect phosphorylated AKT (green) in cardiomyocytes infected with adenovirus Immunofluorescence image of Cryptosporidium parvum oocysts Concentrated groundwater methanotrophic bacteria on 0.2 m filter labeled with fluorescent antibodies....
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This note was uploaded on 12/21/2011 for the course STEP 1 taught by Professor Dr.aslam during the Fall '11 term at Montgomery College.

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Immunofluorescence - Direct immunofluorescence 2. Indirect

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