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Digital Connections. flow cytometer. abhilasha chaware 8202 krutika kuwar 8217.
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Flow cytometer is a laser based instrument which is used in the detection and measurement of physical and chemical characteristics of cells or particles in a heterogeneous fluid mixture..
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The basic principle of flow cytometry is based on the measurement of light scattered by particles, and the fluorescence observed when these particles are passed in a stream through a laser beam..
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FL3 ooo FL2 FLI ssc FSC Amplification Analog-to- Digital Conversion.
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fluidics Optic system Electronics system. Parts of flow cytometry.
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The fluidic system consists of flow cell (quartz chamber).
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The photodetector convert the photons to electrical impulses..
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It converts photons to photoelectrons.. It measures the amplitude of the photoelectrons pulse and converts it into voltage pulse..
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Before running in the flow cytometers, the cells under analysis must be in a single-cell suspension..
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ANTIBODY STAINING:. Once the sample is prepared, the cells are coated with fluoro chrome-conjugated antibodies specific for the surface markers present on different cells. this can be done either by direct, indirect, or intracellular staining..
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TAKE A BIOLOGICAL SAMPLE. BASIC MECHANISM. LABEL IT WITH A FLUORESCENT MARKER.
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Collect and weight Fix with (B) Non—target cell Target cell Antibody detection Mince and digest bilize with rnethanol Stain DNA Live Dead cen Dead cell discrirnination Store at —20 —C Analyze with cytorneter.
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It is used in clinical labs for the detection of malignancy in bodily fluids like leukemia..
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This process doesn’t provide information on the intracellular location or distribution of proteins..
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References. Biotech, M. (2018). Flow cytometry instrumentation – an overview. Current Protocols in Cytometry, e52. DOI: 10.1002/cpcy.52 McKinnon K. M. (2018). Flow Cytometry: An Overview. Current protocols in immunology , 120 , 5.1.1–5.1.11. https://doi.org/10.1002/cpim.40 Dean, P.N. and Hoffman, R.A. (2007), Overview of Flow Cytometry Instrumentation. Current Protocols in Cytometry, 39: 1.1.1-1.1.8. DOI: 1002/0471142956.cy0101s39 https:// enquirebio.com/flow-cytometry.
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Digital Numbers. Thank You.