Clinical Chemistry 46:8(B) 1221–1229 (2000). Flow Cytometry: Principles and Clinical Applications in Hematology. By: Michael Brown and Carl Wittwer
In biotechnology, flow cytometry is a laser- or impedance-based, biophysical technology employed in cell counting, cell sorting, biomarker detection and protein engineering, by suspending cells in a stream of fluid and passing them by an electronic detection apparatus. It allows simultaneous multiparametric analysis of the physical and chemical characteristics of up to thousands of particles per second.
Flow cytometry is routinely used in the diagnosis of health disorders, especially blood cancers, but has many other applications in basic research, clinical practice and clinical trials. A common variation is to physically sort particles based on their properties, so as to purify populations of interest.
Dear Mrs.:
Flow cytometry a cytometric technique in which cells suspended in a fluid flow one at a time through a focus of exciting light, which is scattered in patterns characteristic to the cells and their components; cells are frequently labeled with fluorescent markers so that light is first absorbed and then emitted at altered frequencies. A sensor detecting the scattered or emitted light measures the size and molecular characteristics of individual cells; tens of thousands of cells can be examined per minute and the data gathered is processed by computer.. Best regards
Flow cytometry is a technology that is used to analyse the physical and chemical characteristics of particles in a fluid as it passes through at least one laser. Cell components are fluorescently labelled and then excited by the laser to emit light at varying wavelengths.
The fluorescence can be measured to determine various properties of single particles, which are usually cells. Up to thousands of particles per second can be analysed as they pass through the liquid stream. Examples of the properties measured include the particle’s relative granularity, size and fluorescence intensity as well as its internal complexity. An optical-to-electronic coupling system is used to record the way in which the particle emits fluorescence and scatters incident light from the laser.
Three main systems make up the flow cytometer instrument and these are the fluidics, the optics and the electronics. The purpose of the fluidics system is to transport the particles in a stream of fluid to the laser beam where they are interrogated. Any cell or particle that is 0.2 to 150 μms in size can be analyzed. If the cells are from solid tissue, they require disaggregation before they can be analyzed. Although cells from animals, plants, bacteria, yeast or algae are usually measured, other particles such as chromosomes or nuclei can also be examined. Some particles such as marine algae are naturally fluorescent, but in general, fluorescent labels are required to tag components of the particle. The section of the fluid stream that contains the particles is referred to as the sample core.
The optics system is made up of lasers which illuminate the particles present in the stream as they pass through and scatter light from the laser. Any flourescent molecules that are on the particle emit fluorescence, which is detected by carefully positioned lenses. Generally, the light scattered from up to six or more fluorescences is determined for two different angles. Optical filters and beam splitters then direct the light signals to the relevant detectors, which emit electronic signals proportional to the signals that hit them. Data can then be collected on each particle or event and the characteristics of those events or particles are determined based on their fluorescent and light scattering properties.The electronics system is used to change the light signals detected into electronic pulses that a computer can process. The data can then be studied to ascertain information about a large number of cells over a short period. Information on the heterogeneity and different subsets within cell populations can be identified and measured. Some instruments have a sorting feature in the electronics system that can be used to charge and deflect particles so that certain cell populations can be sorted for further analysis.
The data are usually presented in the form of single parameter histograms or as plots of correlated parameters, which are referred to as cytograms. Cytograms may display data in the from of a dot plot, a contour plot or a density plot
Flow cytometry is a technology that is used to analyse the physical and chemical characteristics of particles in a fluid as it passes through at least one laser. Cell components are fluorescently labelled and then excited by the laser to emit light at varying wavelengths.
The fluorescence can be measured to determine various properties of single particles, which are usually cells. Up to thousands of particles per second can be analysed as they pass through the liquid stream. Examples of the properties measured include the particle’s relative granularity, size and fluorescence intensity as well as its internal complexity. An optical-to-electronic coupling system is used to record the way in which the particle emits fluorescence and scatters incident light from the laser.
Three main systems make up the flow cytometer instrument and these are the fluidics, the optics and the electronics. The purpose of the fluidics system is to transport the particles in a stream of fluid to the laser beam where they are interrogated. Any cell or particle that is 0.2 to 150 μms in size can be analyzed. If the cells are from solid tissue, they require disaggregation before they can be analyzed. Although cells from animals, plants, bacteria, yeast or algae are usually measured, other particles such as chromosomes or nuclei can also be examined. Some particles such as marine algae are naturally fluorescent, but in general, fluorescent labels are required to tag components of the particle. The section of the fluid stream that contains the particles is referred to as the sample core.
The optics system is made up of lasers which illuminate the particles present in the stream as they pass through and scatter light from the laser. Any flourescent molecules that are on the particle emit fluorescence, which is detected by carefully positioned lenses. Generally, the light scattered from up to six or more fluorescences is determined for two different angles. Optical filters and beam splitters then direct the light signals to the relevant detectors, which emit electronic signals proportional to the signals that hit them. Data can then be collected on each particle or event and the characteristics of those events or particles are determined based on their fluorescent and light scattering properties.
flow cytometry is a laser- or impedance-based, biophysical technology employed in cell counting, cell sorting, biomarker detection and protein engineering, by suspending cells in a stream of fluid and passing them by an electronic detection apparatus.
is a technology that is used to analyse the physical and chemical characteristics of particles in a fluid as it passes through at least one laser. Cell components are fluorescently labelled and then excited by the laser to emit light at varying wavelengths.
http://journals.lww.com/amjclinicaloncology/Citation/1991/02000/Flow_Cytometry_and_Sorting,_Second_Edition.26.aspx
http://www.ufjf.br/imunologia/files/2010/03/Flow-Cytometry-introduction.pdf
Modern flow cytometers are able to analyze several thousand particles every second, in "real time," and can actively separate and isolate particles having specified properties. A flow cytometer is similar to a microscope, except that, instead of producing an image of the cell, flow cytometry offers "high-throughput" (for a large number of cells) automatedquantification of set parameters. To analyze solid tissues, a single-cell suspension must first be prepared.
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look in
http://www.ufjf.br/imunologia/files/2010/03/Flow-Cytometry-introduction.pdf
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