Purification of specific cell population by fluorescence activated cell sorting (FACS).

Journal of Visualized Experiments : JoVE
Sreemanti BasuAvijit Ray

Abstract

Experimental and clinical studies often require highly purified cell populations. FACS is a technique of choice to purify cell populations of known phenotype. Other bulk methods of purification include panning, complement depletion and magnetic bead separation. However, FACS has several advantages over other available methods. FACS is the preferred method when very high purity of the desired population is required, when the target cell population expresses a very low level of the identifying marker or when cell populations require separation based on differential marker density. In addition, FACS is the only available purification technique to isolate cells based on internal staining or intracellular protein expression, such as a genetically modified fluorescent protein marker. FACS allows the purification of individual cells based on size, granularity and fluorescence. In order to purify cells of interest, they are first stained with fluorescently-tagged monoclonal antibodies (mAb), which recognize specific surface markers on the desired cell population (1). Negative selection of unstained cells is also possible. FACS purification requires a flow cytometer with sorting capacity and the appropriate software. For FACS, cells in ...Continue Reading

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Citations

Oct 24, 2012·Journal of Immunotherapy·Christopher A KlebanoffNicholas P Restifo
Nov 11, 2011·Journal of Biomechanical Engineering·Robert KissNicholas A Willoughby
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Feb 7, 2019·Gastric Cancer : Official Journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association·Jong Won LeeYeul Hong Kim
Dec 22, 2017·Nature Protocols·Takamasa KudoMarkus W Covert
Jan 10, 2020·Lab on a Chip·Ewa GuzniczakHelen Bridle
Jun 10, 2017·Clinical and Translational Medicine·Raquel Cuevas-Diaz DuranJia Qian Wu
Sep 2, 2015·The Journal of Immunology : Official Journal of the American Association of Immunologists·Avijit RayBonnie N Dittel
Sep 16, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Thaddäus StrukHermann Pavenstädt
Oct 14, 2017·Scientific Reports·Trisha M WesterhofEdward L Nelson
Aug 9, 2020·Scientific Reports·James D HernandezEleanna De Filippis
Apr 18, 2018·Apoptosis : an International Journal on Programmed Cell Death·Jingwen FengYuanming Feng
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Mar 18, 2021·Journal of Neural Engineering·Aundrea F BartleyLynn E Dobrunz
Jul 3, 2021·Frontiers in Immunology·Eriberto N NataliEnkelejda Miho

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Methods Mentioned

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