lec17 Cytometry

lec17 Cytometry - EE 480/580 Biomedical Electronics Lecture...

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1 Biomedical Electronics Lecture 18 EE 480/580
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2 b Flow cytometry Today Flow Cytometry is the technological process that allows for the individual measurements of cell fluorescence and light scattering. This process is performed at rates of thousands of cells per second. This information can be used to individually sort or separate subpopulations of cells.
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3 b Imagine it to be a lot like visiting a supermarket b You choose the goods and take them to the cashier b Usually you have to pile them onto a conveyor b The clerk picks up one item at a time and interrogates it with a laser to read the barcode b Once identified and if sense prevails, similar goods are collected together, e.g. fruit and vegetables go into one shopping bag and household goods into another b Now picture in your mind the whole process automated; replace shopping with biological cells; and substitute the barcode with cellular markers – this is flow cytometry and cell sorting! Flow cytometry in simple terms
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4 • Flow cytometry developed from microscopy. Thus Leeuwenhoek is often cited in any discussion regarding it’s history. • F.T. Gucker (1947)build the first apparatus for detecting bacteria in a laminar sheath stream of air. • L. Kamentsky (IBM Labs), and M. Fulwyler (Los Alamos Nat. Lab.) experimented with fluidic switching and electrostatic cell sorters respectively. Both described cell sorters in 1965. History
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5 • M. Fulwyler utilized Pulse Height Analyzers to accumulate distributions from a Coulter counter. This feature allowed him to apply statistical analysis to samples analyzed by flow. • In 1972 L. Herzenberg (Stanford Univ.), developed a cell sorter that separated cells stained with fluorescent antibodies.The Herzenberg group coined the term Fluorescence Activated Cell Sorter (FACS). History
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6 Fluorescence Activation Process (or Immunofluorescence) FITC FITC FITC FITC Antibodies recognize specific molecules in the surface of some cells but not others When the cells are analyzed by flow cytometry the cells expressing the marker for which the antibody is specific will manifest fluorescence. Cells who lack the marker will not manifest fluorescence Antibodies are artificially conjugated to fluorochromes Antibodies
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7 Flow Cytometry Applications • Immunofluorescence • Cell Cycle Kinetics • Cell Kinetics • Genetics • Molecular Biology • Microbiology • Biological Oceanography • Parasitology • Bioterrorism
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EE 480/580 Fall 2010 8 • Flow cytometry integrates electronics, fluidics, computer, optics, software, and laser technologies in a single platform Flow cytometry
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9 b Flow cytometry is a technique for the analysis of multiple parameters of individual cells within heterogeneous populations. b
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lec17 Cytometry - EE 480/580 Biomedical Electronics Lecture...

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