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Related Concept Videos

Protein Dynamics in Living Cells01:19

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Related Experiment Video

Updated: Jul 8, 2025

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

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Real-time open-source FLIM analysis.

Kevin K D Tan1,2, Mark A Tsuchida2, Jenu V Chacko2

  • 1Department of Biomedical Engineering, University of Wisconsin, Madison, WI, United States.

Frontiers in Bioinformatics
|December 15, 2023
PubMed
Summary
This summary is machine-generated.

We developed the first open-source real-time analysis tool for Fluorescence Lifetime Imaging Microscopy (FLIM) data. This tool enables on-the-fly quality assessment during imaging, reducing session times and sample damage.

Keywords:
FLIMfluorescence lifetimelive FLIMnapariopen sourcephasorreal-time FLIM analysis

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Area of Science:

  • Microscopy
  • Biophotonics
  • Data Analysis

Background:

  • Fluorescence Lifetime Imaging Microscopy (FLIM) offers quantitative insights into fluorophore microenvironments.
  • Traditional FLIM analysis, especially with time-correlated single-photon counting (TCSPC), is time-consuming and performed post-acquisition.
  • This delay leads to data quality uncertainties, extended imaging times, and potential photobleaching or photodamage.

Purpose of the Study:

  • To introduce the first open-source program for real-time FLIM data analysis during specimen scanning.
  • To enable on-the-fly assessment of FLIM data quality, improving efficiency and reducing risks to sensitive samples.
  • To integrate real-time computational and visualization capabilities with existing acquisition software.

Main Methods:

  • Developed an open-source real-time FLIM viewer as a Napari plugin.
  • Integrated real-time data transmission from acquisition software (e.g., OpenScan).
  • Implemented phasor analysis and rapid lifetime determination (RLD) for immediate results.

Main Results:

  • The tool provides real-time FLIM data quality assessment during acquisition.
  • Facilitates early identification of FLIM signatures and potential issues.
  • Significantly speeds up the imaging process, especially for live biological samples.

Conclusions:

  • This real-time analysis approach enhances the efficiency and reliability of FLIM experiments.
  • It minimizes the need for extended imaging sessions, preserving sample integrity.
  • The open-source nature promotes accessibility and further development in quantitative FLIM.