FLOW ESSAY
Lynnzie Tompkins
December 6, 2014
Dr. Jutila
Flow Cytometry (BIOB413)
A look into Flow Cytometry
Flow cytometry is a widely used practice in the biotechnology field; it is mainly used routinely for the diagnosis of health disorders. However, flow cytometry is also commonly used, as an application for basic research. Simply, flow cytometers are automated machines that determine properties of individual cells (e.g. cell size, cell granularity, and total DNA). Cytometers are similar to microscopes, except that, instead of creating an image of the cell, the flow cytometer has the ability to analyze and sort a large amount of cells in real time using set parameters that are determined by the user. The cytometer contains a laser-based technology that counts and sort’s cells by suspending them in a fluid stream, each cell is then passed over a electronic detector. The detector allows for synchronic analysis of both the physical (e.g. cell size, cell granularity), and chemical (e.g. total DNA/RNA) properties of the particles with in the sample being analyzed.
A traditional flow cytometer contains three main components: optics, fluidics, and electronics. For the optics of the cytometer, most commonly a laser beam is used as the light source. To be more specific an argon laser, which produces blue light is most commonly used however, some instruments also contain a red helium-neon laser. The laser beam excites a fluorescent tag, for example a fluorescent antibody, which is bound onto the cells that are within the sample. Before the laser excites the tag it is focused through optics, which form an elliptical beam that contains its maximal intensity in the center. Once the laser strikes the cell the light is diffracted around the cells edges. The scattered light is approximately equal to the circumference of the cell. Hence the term forward scatter (FSC). The light from the laser that is reflected off of the cell and its internal structures is described as the
December 6, 2014
Dr. Jutila
Flow Cytometry (BIOB413)
A look into Flow Cytometry
Flow cytometry is a widely used practice in the biotechnology field; it is mainly used routinely for the diagnosis of health disorders. However, flow cytometry is also commonly used, as an application for basic research. Simply, flow cytometers are automated machines that determine properties of individual cells (e.g. cell size, cell granularity, and total DNA). Cytometers are similar to microscopes, except that, instead of creating an image of the cell, the flow cytometer has the ability to analyze and sort a large amount of cells in real time using set parameters that are determined by the user. The cytometer contains a laser-based technology that counts and sort’s cells by suspending them in a fluid stream, each cell is then passed over a electronic detector. The detector allows for synchronic analysis of both the physical (e.g. cell size, cell granularity), and chemical (e.g. total DNA/RNA) properties of the particles with in the sample being analyzed.
A traditional flow cytometer contains three main components: optics, fluidics, and electronics. For the optics of the cytometer, most commonly a laser beam is used as the light source. To be more specific an argon laser, which produces blue light is most commonly used however, some instruments also contain a red helium-neon laser. The laser beam excites a fluorescent tag, for example a fluorescent antibody, which is bound onto the cells that are within the sample. Before the laser excites the tag it is focused through optics, which form an elliptical beam that contains its maximal intensity in the center. Once the laser strikes the cell the light is diffracted around the cells edges. The scattered light is approximately equal to the circumference of the cell. Hence the term forward scatter (FSC). The light from the laser that is reflected off of the cell and its internal structures is described as the
Cited: Analysis Of Cell Cycle By Flow Cytometry, and 301. Analysis of Cell Cycle by Flow Cytometry (n.d.): n. pag. Web. 05 Dec. 2014. Bakke, Phd Antony C. "The Principles of Flow Cytometry." Laboratory Medicine 32.4 (2001): 207-11. Web. 05 Dec. 2014. "Cell-cycle Analysis." Wikipedia. Wikimedia Foundation, 12 Feb. 2014. Web. 05 Dec. 2014. "Flow Cytometric Analysis of Cell Cycle with Propidium Iodide DNA Staining." Flow Cytometric Analysis of Cell Cycle with Propidium Iodide. N.p., n.d. Web. 05 Dec. 2014. "Introduction to Flow Cytometry." Introduction to Flow Cytometry. N.p., n.d. Web. 05 Dec. 2014. "What Is Flow Cytometry?" What Is Flow Cytometry? N.p., n.d. Web. 05 Dec. 2014.