8/14/2023 0 Comments Define flow cytometry![]() Cell sorting- A particular subset(s) of cells may be sorted from a mixture of cells based upon particular properties.Cell proliferation- Carboxyfluorescein diacetate, succinimidyl ester (CFSE) is a dye that diffuses into cells and is passed from parent to daughter cells so that the cells of each generation have half the fluorescent intensity of their parent cells.two copies of a genome have roughly twice the fluorescent intensity of one copy). Fluorescent intensity is used to determine the amount of cellular DNA present in each cell (i.e.Cell cycle analysis- Fixed cells are stained with a dye that binds to DNA (e.g.Cells can be stained with Annexin V or 7ADD. Apoptosis measurement- Several flow cytometric methods to detect apoptosis are available.Transfection efficiency may be determined when a fluorescent protein (i.e.Immunophenotyping- Using fluorescence-conjugated antibodies directed toward a protein(s) of interest, cells expressing that protein(s) on the surface or intracellularly may be detected by flow cytometry.Specific cell types may be distinguished within a mixed population using multiple fluorescence-conjugated antibodies.There are a number of applications for flow cytometry, including, but not limited to: The cells or particles can be separated by sorting, or the information can be collected and analyzed. The result is a visual presentation describing an individual or group of cellular events. Information about the size and granularity of a cell is recorded, as well. The fluorescent signal is detected and amplified, then translated into an electronic signal, which is sent to the computer. The fluorescent probes are excited by the laser and then emit light. ![]() The cells then pass by a laser, which emits a specific wavelength of light. Cells or particles tagged with fluorescent molecules enter the cytometer via a fluid stream. Flow cytometry uses fluorescent probes to identify and characterize cells or particles. The benefit of flow cytometry is the rapid simultaneous measurement of several parameters on a cell by cell basis. Flow Cytometry is a means of identifying and measuring certain physical and chemical characteristics of cells or particles as they travel in suspension. ![]()
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