Flow Cytometry: Basics and Applications
Pp. 27-42 (16)
Juhi Saxena and Anupam Jyoti
Flow cytometer is a sophisticated and analytical tool which analyzes the
phenotypic and functional characteristics of a cell in much lesser time. It measures the
light scattering features, mainly forward scatter and side scatter properties of the cell.
Optics, fluidics, and electronics are the main elements flow cytometer. Hydrodynamic
focusing of the sheath fluid present in a sheath chamber enables the streamline motion
of cells. This allows the encounter of each cell separately with the laser. A photodiode,
photomultiplier tubes, optical filters, and beam splitters collect and emit the light or
fluorescence in the form of signals. The signals are further processed by the electronic
component and with the help of software, data are analyzed. For a cell to emit
fluorescence, a number of fluorochromes (FITC, Alexa Fluor 488, Alexa Fluor 647
etc.) and dyes (propidium iodide, Hoechst, ethidium bromide etc.) are available. These
fluorochromes may be conjugated with antibodies and emit fluorescence when excited
by the laser. A diverse range of applications are possible using flow cytometer like
apoptosis assay, cell cycle analysis, free radical generation assay, cytokine estimation,
immunophenotyping, phagocytosis assay and many more. An advanced application of
flow cytometer receiving attention is cell sorting. Individual cells or cell types can be
sorted from a mixture of cells using physical fluorescence emission features of the cells
named fluorescence assisted cell sorter.
Apoptosis, Cell cycle, Cells sorting, Fluorochromes, Fluorescence,
Immunophenotyping, Laser, Light scattering, Photomultiplier tube.
Amity Institute of Biotechnology, Amity University Rajasthan, Amity Education Valley, Kant Kalwar, NH-11C, Jaipur-Delhi Highway, Jaipur, India.