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Single-Cell Characterization of Calcium Influx and HIV-1 Infection using a Multiparameter Optofluidic Platform
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1Department of Medicine, Division of Infectious Diseases,Icahn School of Medicine at Mount Sinai, 2Department of Genetics and Genomic Sciences,Icahn School of Medicine at Mount Sinai, Icahn Institute of Data Science and Genomic Technology, 3Black Family Stem Cell Institute, Department of Genetics and Genomic Sciences,Icahn School of Medicine at Mount Sinai, Icahn Institute of Data Science and Genomic Technology, 4Sema4, a Mount Sinai venture
Chapters
- 00:05Introduction
- 00:57Preparation of Cells for Imaging
- 02:33Optofluidic System Preparation, Cell Loading, and Cell Penning
- 04:27Single-cell Infection and Functional Imaging
- 05:24Results: Single-Cell Characterization of Calcium Influx and HIV-1 Infection
- 06:20Conclusion
Here, we present a protocol in which single cells are monitored for acute events and productive HIV-1 infection on a nanofluidic device. Imaging data define virus-host receptor interactions and signaling pathway dynamics. This is the first method for nanofluidic high-throughput longitudinal single-cell culture and imaging to study signaling kinetics and molecular interactions.
