Fluorescence–activated cell sorting

Fluorescence–activated cell sorting
• Fluorescence–activated cell sorting (FACS) was invented to sort a
heterogeneous mixture of cells into different homogenous
subpopulations of interest based upon the specific light scattering
and fluorescent characteristics of each cell. In a complex cell mixture,
the different homogenous subpopulations may have different
antigenic and other markers on their surface. These markers can be
tagged by the means of fluorescent labels which can be detected by
laser and light detectors of fluorescence-activated cell sorting. It
provides rapid, accurate and high throughput cell analysis and has
been applied to examine diverse biological processes such as cell
cycle, cell proliferation, cell viability, cell phenotyping, cell signaling,
micronucleus test, intracellular cytokine secretion.

• Fluorescence–activated cell sorting determine cells automatically based either
on cellular properties or by fluorescent labeling. The ability to sort cells based
on physical characteristic and their fluorescent label signatures enables to
isolate well-defined subpopulations of cells in more effective manner than
other separation methods likely magnetic separation. The use of fluorescent
colors facilitates to detect simultaneously different subpopulations of interest,
it is time-effective and labor-saving. Green fluorescent protein (GFP) is used as
a reporter gene. another complementary is monoclonal antibodies, which is
highly specific for its target antigens and can readily be coupled with
fluorescein, phycobiliproteins and other fluorochromes. Monoclonal
antibodies widely used as FACS reagents enhanced the definition of hundreds
of target antigens present on or in cells. FACS is also the most efficient method
of cloning cells, especially when they are present in a extremely low frequency.

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