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URL: https://dx.doi.org/DOI:10.17504/protocols.io.mgdc3s6

Authors: Cj Xia

Group: Boster Bio

Summary: Fluorescent activated cell sorting (FACS) is a specialized type of flow cytometry used for sorting and analyzing a heterogeneous mixture of cells into different subpopulations based on the specific light scattering and fluorescent characteristics (from the specific labels) of each cell. The number of measurable parameters that can be used by this technology to separate cell populations is immense – starting from simple surface immunophenotyping to metabolic functions, cell cycle status, redox state, and DNA content analysis to name a few. Since its inception, FACS has been used extensively in biomedical research and clinical diagnostics and therapeutics. The most common usage of FACS is seen in: - Analysis of whole human blood for diagnosing diseases, immunophenotyping - Sorting different blood cell fractions for ex-vivo manipulations and/or transplantations - Immuno-phenotypic analysis of murine blood to identify transgenic/knockout animals - Sorting and analysis of a slew of cell lines for various biological assays - Characterization and isolation of rare cells types like adult stem cells and cancer initiating cells Each human cell expresses hundreds of thousands of cell surface antigens that specify their cell type, biological function, development stage, and much more. Cells residing in different organs have characteristic cell surface antigens, and determination of these cells using the specific fluorophore-conjugated antibodies can be analyzed by flow cytometry. The following general protocols are recommended for various common FACS staining procedures. Staining with unconjugated purified antibody needs an additional step of staining with a fluorescent conjugated secondary antibody (indirect immunostaining).

Affiliations: University of Virginia, Charlottesville

External URL: https://www.bosterbio.com/protocol-and-troubleshooting/flow-cytometry-protocol

Version: 1

Publication Date: 2018

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