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Quantitative High-throughput Single-cell Cytotoxicity Assay For T Cells
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Quantitative High-throughput Single-cell Cytotoxicity Assay For T Cells
Capitoli
- 00:05Titolo
- 01:32Fabrication of Arrays of Nanowells in PDMS
- 02:14Target Cell (T) Labeling
- 02:53Effector (E) Cell Labeling
- 03:26Separation of Live and Dead Cells by Ficoll Density Gradient
- 05:23Cell Loading onto Nanowell Array
- 06:51Imaging
- 07:58Results: A High-throughput Cytolytic Assay
- 09:03Conclusion
We describe a single-cell high-throughput assay to measure cytotoxicity of T cells when incubated with tumor target cells. This method employs a dense, elastomeric array of sub-nanoliter wells (~100,000 wells/array) to spatially confine the T cells and target cells at defined ratios and is coupled to fluorescence microscopy to monitor effector-target conjugation and subsequent apoptosis.
