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Live-cell Imaging of Single-Cell Arrays (LISCA) – a Versatile Technique to Quantify Cellular Kinetics
JoVE Journal
Biologia
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JoVE Journal Biologia
Live-cell Imaging of Single-Cell Arrays (LISCA) – a Versatile Technique to Quantify Cellular Kinetics
DOI:

10:24 min

March 18, 2021

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Capítulos

  • 00:04Introduction
  • 00:30Microstructured Single Cell Array Fabrication
  • 03:16Cell Seeding
  • 04:21Perfusion System
  • 05:13Time-Lapse Microscopy
  • 06:44Fluorescent Marker mRNA Transfection
  • 07:34Image Analysis and Fluorescence Readout
  • 08:29Results: Representative Image Analysis and Fluorescence Readout
  • 09:34Conclusion

Summary

Tadução automática

We present a method for the acquisition of fluorescence reporter time courses from single cells using micropatterned arrays. The protocol describes the preparation of single-cell arrays, the setup and operation of live-cell scanning time-lapse microscopy and an open-source image analysis tool for automated preselection, visual control and tracking of cell-integrated fluorescence time courses per adhesion site.

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