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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Live-cell Imaging of Single-Cell Arrays (LISCA) - a Versatile Technique to Quantify Cellular Kinetics
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Published on: March 18, 2021

Quantitative time-lapse fluorescence microscopy in single cells.

Dale Muzzey1, Alexander van Oudenaarden

  • 1Department of Physics, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.

Annual Review of Cell and Developmental Biology
|July 7, 2009
PubMed
Summary
This summary is machine-generated.

Green fluorescent protein (GFP) enables cell biology visualization. Combined with computational image analysis, time-lapse microscopy now offers quantitative, high-throughput insights into dynamic cellular processes and organism development.

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Live-Cell Fluorescence Microscopy to Investigate Subcellular Protein Localization and Cell Morphology Changes in Bacteria

Published on: November 23, 2019

Area of Science:

  • Cell Biology
  • Microscopy
  • Computational Biology

Background:

  • Green fluorescent protein (GFP) revolutionized cell biology by enabling visualization of molecular mechanisms in living cells.
  • Fluorescence microscopy has advanced from qualitative assessments to highly quantitative and high-throughput analyses.
  • Computational image analysis is crucial for processing large datasets generated by modern microscopy.

Purpose of the Study:

  • To review studies utilizing time-lapse fluorescence microscopy and automated image analysis for single-cell dynamic event investigation.
  • To highlight how single-cell analysis provides unique mechanistic insights beyond bulk assays.
  • To discuss advancements in imaging technology for simultaneous perturbation and monitoring of biological systems.

Main Methods:

  • Time-lapse fluorescence microscopy
  • Automated computational image analysis
  • Single-cell level analysis
  • 4D lineage tracing in developing organisms

Main Results:

  • Single-cell analysis offers unique mechanistic insights into cellular processes.
  • Quantitative microscopy facilitates detailed 4D lineage assembly in developing organisms.
  • Recent advances include platforms for simultaneous perturbation and quantitative monitoring.

Conclusions:

  • The integration of fluorescence microscopy and computational analysis has transformed cell biology research.
  • Quantitative single-cell approaches provide deeper understanding of dynamic biological systems.
  • Emerging imaging technologies promise further advancements in biological system analysis.