Flow Cytometer

Flow Cytometer

Janice Dominov, Ph.D., Scientist

 

Flow cytometry (FCM) is a method for quantifying the types of cells in a mixture.  It may be used simply for cell analysis or it can be used for cell sorting, where the subpopulations are physically separated from each other and used for further study.  No other method allows as rapid a quantitative and detailed analysis of subpopulations of cells as FCM.  It is popular for both research and clinical diagnosis, because it can quickly, accurately and inexpensively analyze a large number of samples (30-50 samples/hour).

 

No other method allows as a rapid a quantitative and detailed analysis of subpopulations of cells as flow cytometry.

To do the analysis, fluorescently labeled cells in solution are passed single-file as a fine stream through a laser beam.  Each cell scatters some of the laser light and also emits fluorescent light excited by the laser. The cytometer typically measures several parameters simultaneously for each cell: a) the low-angle forward scatter intensity, approximately proportional to the cell diameter; b) the orthogonal (at 90º angle) scatter intensity, approximately proportional to the quantity of granular structures within the cell; and c) fluorescence intensities at 3-4 wavelengths. The computer records the data and displays them graphically.

 

In research, fluorescent antibodies are often used to measure specific cell components such as surface receptors, expression of a transgene, or binding of viruses or hormones to their receptors. Another typical measurement is the amount of total DNA per cell for analyzing the growth and death of cells. FCM also has important clinical applications in diagnosis, prognosis, and optimization of therapy. For example, FCM can distinguish subtle changes in the types of blood cells to discriminate between different types of blood cell cancers (e.g. leukemias and lymphomas).

 

FACS, but no Fax.  BBRI owns a flow cytometry instrument called FACSCalibur that can perform both cell analysis and cell sorting. Flow cytometry is a generic term and is abbreviated as FCM, while FACS (Fluorescence Activated Cell Sorter) is a trademark. FACS is pronounced just like Fax.  FACS has been around since the 1960s, whereas facsimile transmission arose only in the 1980s. Therefore, flow cytometrists see no need to stop speaking of "the FACS".