Flow Cytometry

Flow Cytometry measures a variety of parameters of cells within heterogenous populations. It can measure phenotypes, density of cell surface molecules, cell count etc.
Cells that are to be analyzed in this case for detection of certain antigens/molecules on cell surfaces are washed with a saline solution and introduced to a mouse antibody directed against the antigen(s) of interest. If the matching antigen exist on the cell surface, the antibody sticks to it. The cells with a possible antibody attached to a particular antigen is washed again with a saline solution (PBS) to get rid of excess antibodies that did not stick. The cells are then introduced to a goat anti-mouse antibody, which if the mouse antibody is present (attached to the antigen of interest), than the goat anti-mouse antibody will stick the the mouse antibody. The goat anti-mouse antibody is tagged with a particular fluoraphore (a molecule that fluoresces a particular color of light). Then, these cells are passed through a beam of light which excites the fluoraphore and then allows it to release energy with a particular wavelengh of color. The color is then detected by the flow cytometer machine and is sent to a computer, where the computer analyses the data and shows if the antigen of interest was present on the cell, and how much of it was present
*This is one of many uses of flow cytometry

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