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Immunofluorescence (IF) is a light microscopy-based technique that allows detection and localization of a wide variety of target biomolecules within a cell or tissue at a quantitative level. The technique utilizes the binding specificity of antibodies and antigens . [ 1 ]
Immune electron microscopy (more often called immunoelectron microscopy) is the equivalent of immunofluorescence, but it uses electron microscopy rather than light microscopy. [1] Immunoelectron microscopy identifies and localizes a molecule of interest, specifically a protein of interest, by attaching it to a particular antibody .
Immunocytochemistry is a technique used to assess the presence of a specific protein or antigen in cells (cultured cells, cell suspensions) by use of a specific antibody, which binds to it, thereby allowing visualization and examination under a microscope. It is a valuable tool for the determination of cellular contents from individual cells.
Immunogold labeling was first used in 1971 by Faulk and Taylor to identify Salmonella antigens. [2] [4] It was first applied in transmission electron microscopy (TEM) and was especially useful in highlighting proteins found in low densities, such as some cell surface antigens. [5]
Direct FA stained mouse brain impression smear reveals the presence of the bacterium Chlamydia psittaci. 400X.. A direct fluorescent antibody (DFA or dFA), also known as "direct immunofluorescence", [1] is an antibody that has been tagged in a direct fluorescent antibody test.
Electron microscopy or EM can be used to study the detailed microarchitecture of tissues or cells. Immuno-EM allows the detection of specific proteins in ultrathin tissue sections. Antibodies labelled with heavy metal particles (e.g. gold) can be directly visualised using transmission electron microscopy. While powerful in detecting the sub ...
Colocalization is used in real-time single-molecule fluorescence microscopy to detect interactions between fluorescently labeled molecular species. In this case, one species (e.g. a DNA molecule) is typically immobilized on the imaging surface, and the other species (e.g. a DNA-binding protein) is supplied to the solution.
Immunofluorescence coloration of actin (green) and the focal adhesion protein vinculin (red) in a fibroblast. Focal adhesions are visible as red dots at the end of the green bundles. In cell biology , focal adhesions (also cell–matrix adhesions or FAs ) are large macromolecular assemblies through which mechanical force and regulatory signals ...