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Flow cytometry (FC) is a technique used to detect and measure the physical and chemical characteristics of a population of cells or particles. [1] [2] [3] [4]In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument.
Cell cycle analysis by DNA content measurement is a method that most frequently employs flow cytometry to distinguish cells in different phases of the cell cycle.Before analysis, the cells are usually permeabilised and treated with a fluorescent dye that stains DNA quantitatively, such as propidium iodide (PI) or 4,6-diamidino-2-phenylindole (DAPI).
Schematic diagram of a flow cytometer, showing focusing of the fluid sheath, laser, optics (in simplified form, omitting focusing), photomultiplier tubes (PMTs), analogue-to-digital converter, and analysis workstation. Flow cytometers operate by hydrodynamically focusing suspended cells so that they separate from each other within a fluid stream.
Flow cytometry cell-sorting yields very high specificity according to one or several surface markers, but one limitation is constituted by the number of cells that can be processed during a work day. For this reason pre-enrichment of the population of interest by immunomagnetic cell sorting is often considered, especially when the target cells ...
Flow cytometry is a method in cell biology that employs the deflection of laser light a well as the excitation of fluorescent dyes to analyse various properties of a high number cells in a relatively short time. This category lists methods and tools used in flow cytometry.
A wide (hundreds of micrometers in diameter) tube made of glass or plastic is used, through which a "wall" of fluid called the sheath flow is pumped. The sample is injected into the middle of the sheath flow. If the two fluids differ enough in their velocity or density, they do not mix: they form a two-layer stable flow. [2]
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Flow cytometry is by far the most sophisticated and expensive method for cell counting. In a flow cytometer the cells flow in a narrow stream in front of a laser beam. The beam hits them one by one, and a light detector picks up the light that is reflected from the cells.