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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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Related Experiment Video

Updated: Jun 2, 2026

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
09:30

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

Published on: January 18, 2017

AlliGator: Open source fluorescence lifetime imaging analysis in G.

Xavier Michalet1

  • 1Department of Chemistry & Biochemistry, University of California at Los Angeles, 607 Charles E. Young Drive E, Los Angeles, CA 90095, United States of America.

Softwarex
|June 1, 2026
PubMed
Summary
This summary is machine-generated.

AlliGator is a new open-source software for analyzing Fluorescence Lifetime Imaging (FLI) data. It offers user-friendly tools for complex analyses like Förster Resonant Energy Transfer (FRET) and membrane potential mapping.

Keywords:
Fluorescence lifetime imagingLabVIEWNLSFPhasor analysisPython

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Last Updated: Jun 2, 2026

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Flexible Measurement of Bioluminescent Reporters Using an Automated Longitudinal Luciferase Imaging Gas- and Temperature-optimized Recorder (ALLIGATOR)

Published on: December 13, 2017

Area of Science:

  • Biophysics
  • Optical Imaging
  • Software Development

Background:

  • Fluorescence Lifetime Imaging (FLI) captures temporal fluorescence decay at each pixel.
  • FLI data analysis typically involves complex model fitting or phasor analysis.
  • Existing software may lack user-friendliness or specific analytical features.

Purpose of the Study:

  • To introduce AlliGator, a novel open-source software for FLI data analysis.
  • To provide a user-friendly graphical interface for advanced FLI techniques.
  • To enable extended functionality through Python-based plugins.

Main Methods:

  • AlliGator implements nonlinear least-square fit and maximum likelihood estimation for decay analysis.
  • Phasor analysis is incorporated for alternative FLI data interpretation.
  • The software utilizes LabVIEW for its graphical user interface (GUI) and supports Python plugins.

Main Results:

  • AlliGator offers a user-friendly GUI for FLI data analysis.
  • Unique features include Förster Resonant Energy Transfer (FRET) stoichiometry analysis.
  • The software can generate maps of lifetime-derived quantities, such as membrane potential.

Conclusions:

  • AlliGator provides a versatile and accessible platform for FLI data analysis.
  • Its open-source nature and plugin support foster extensibility.
  • The software facilitates advanced biophysical and imaging research.