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Journal / Immunology and Infection / "Phagosome Closure Assay" to Visualize Phagosome…
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Immunology and Infection

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JoVE Journal Immunology and Infection
"Phagosome Closure Assay" to Visualize Phagosome Formation in Three Dimensions Using Total Internal Reflection Fluorescent Microscopy (TIRFM)
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"Phagosome Closure Assay" to Visualize Phagosome Formation in Three Dimensions Using Total Internal Reflection Fluorescent Microscopy (TIRFM)

Article DOI: 10.3791/54470-v • 10:07 min • August 26th, 2016
August 26th, 2016 •
1, 1, 1, 1
1Inserm U1016, Institut Cochin, CNRS UMR 8104, Université Paris Descartes, Sorbonne Paris Cité

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Summary
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We describe an experimental setup to visualize with unprecedented high resolution phagosome formation and closure in three dimensions in living macrophages, using total internal reflection fluorescence microscopy. It allows monitoring of the base of the phagocytic cup, the extending pseudopods, as well as the precise site of phagosome scission.

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Phagosome Closure Assay Visualize Phagosome Formation TIRF Microscopy Total Internal Reflection Fluorescent Microscopy Macrophages Pseudopod Extension Tip Fusion Fluorescent Proteins Critical Angle Plasma Membrane Heating Chamber Sheep Red Blood Cells (SRBCs) Glass-bottom Dish PBS BSA Rabbit IGG Anti-SRBCs Incubation Serum-free Microscopy Medium
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