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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.
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.
The whole procedure can be performed on cells from the blood, bone marrow or spinal fluid in a matter of a few hours. [citation needed] Immunophenotyping is a very common flow cytometry test in which fluorophore-conjugated antibodies are used as probes for staining target cells with high avidity and affinity.
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).
Further information about the specimen may be gained by immunohistochemical stains and molecular testing, particularly if the sample is prepared using liquid based cytology. Often "reflex" testing is performed, such as HPV testing on an abnormal pap test or flow cytometry on a lymphoma specimen.
High-content screening technology is mainly based on automated digital microscopy and flow cytometry, in combination with IT-systems for the analysis and storage of the data. "High-content" or visual biology technology has two purposes, first to acquire spatially or temporally resolved information on an event and second to automatically ...
This file contains the total ion counts for each channel for every cell arranged in a matrix and is the same file generated during flow cytometry. [5] Manual gating of this data can be performed as is done for flow cytometry and most of the tools available for flow cytometry analysis have been ported to CyTOF (See flow cytometry bioinformatics ...
However, flow cytometry can be less effective at detecting extremely rare cell populations, and there is a loss of architectural relationships in the absence of a tissue section. [5] Flow cytometry also has a high capital cost associated with the purchase of a flow cytometer. [citation needed]