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“Phagosome Closure Assay” to Visualize Phagosome Formation in Three Dimensions Using Total Internal Reflection Fluorescent Microscopy (TIRFM)
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Capítulos
- 00:05Título
- 00:51Red Blood Cell (RBC) Opsonization and Poly-lysine Coverslip Coating
- 02:31Non-covalent SRBC Fixation
- 04:25TIRF Microscopy Phagocytosis Visualization
- 05:23Image Acquisition and Processing
- 07:45Results: F-actin Accumulation at the Site of Phagosome Closure
- 08:51Conclusion
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.
