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Background on Flow Cytometry
Flow cytometry
(FCM) is a technology that allows cells in suspension
to be efficiently measured at
rates of up to 10,000 cells/second. As cells pass through a
focused laser beam, the cell size, fluorescent dye distribution,
internal structure, membrane potential, nuclear diameter may be determined, as well as, the detection
DNA, enzymes, RNA, and surface antigens. Flow Cytometry and the Micronucleus Test Traditionally, the scoring of micronuclei (MN) was performed by microscopic inspection of 1,000 to 2,000 young Red Blood Cells (reticulocytes). This was, by necessity, a very subjective, time- and labor-intensive method, giving wide standard deviations, and a greater chance for technician error. In contrast, flow cytometers offer speeds far beyond that
of microscopic inspection or image analysis. With this increased
speed and objectivity, it is now possible to score 20,000 reticulocytes
for micronuclei, ten times the data, in just minutes. Rather than waiting
weeks or months for results, a researcher can obtain data from a typical test in just
one day. This is an enormous benefit to those researchers working
in competitive fields. Flow cytometric analysis
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The results from flow cytometric analysis of a typical blood sample are
shown at right. The Y axis corresponds to green fluorescence (FL1) and the
X axis to red fluorescence (FL3). When analyzing micronucleated cells,
the DNA (micronuclei) of each cell is stained with a red dye, while
the RETs (youngest cells) are stained green.