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What is Flow Cytometry? |
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Flow cytometry is a technology that measures and analyzes the optical properties of mono-dispersed single particles, such as cells, bacteria, picoplankton, microbeads, yeast, platelets, nuclei and other similarly-sized particles, passing single file through a focused laser beam. The laser can excite fluorophores that have been used to mark various molecules or physiological functions of the particles. The use of fluorophores with different fluorescence characteristics, multiple lasers and multiple photo detectors allows flow cytometers to measure many characteristics of each particle simultaneously. An important feature of flow cytometry is that large numbers, for example thousands of particles per second, are analyzed and therefore provide a statistically significant picture of a specimen's physical and biochemical make-up. To perform these functions, cytometers requires a combined system of:
A further strength of flow cytometry, unlike fluorescent microscopy, is that it is able to distinguish between multiple closely emitting fluorophores, such as eGFP and eYFP, in concurrently expressing cells if the appropriate optics and experimental controls exist. If sort criteria have been set up, some flow cytometers can physically sort selected sub-populations. Optical signals consisting of laser light scatter and fluorescence are sequentially generated by each single particle and consist of the following:
Common applications of flow cytometry include the analysis and/or sorting of experiments studying:
There is a nice overview article of flow cytometry in Nature Methods. A valuable forum for the discussion of many aspects of flow cytometry is the Cytometry Electronic Mailing List. To subscribe, contact cyto-request@flowcyt.cyto.purdue.edu. |
Jinghang Zhang is solely responsible for the content of this site. Comments, concerns and questions regarding it should be addressed to her. |