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Updated: Jun 2, 2026

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
1Department of Chemistry & Biochemistry, University of California at Los Angeles, 607 Charles E. Young Drive E, Los Angeles, CA 90095, United States of America.
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Fluorescence Lifetime Imaging (FLI) is a technique recording the temporal decay of fluorescence emission at every pixel of an image. Analyzing the information embedded in FLI dataset requires either fitting the decay to a predefined model using nonlinear least-square fit or maximum likelihood estimation, or projecting the decay on an orthogonal basis of periodic functions as in phasor analysis. AlliGator is a Windows open source software (BSD license) implementing these approaches in a user-friendly graphical user interface (GUI) and offering numerous unique features such as Förster Resonant Energy Transfer (FRET) stoichiometry analysis, or the creation of maps of lifetime-derived quantities such as membrane potential. It leverages the unique ability of the LabVIEW graphical programming language (G) to design feature-rich GUI, and supports user-developed plugins written in python to extend its native capabilities.

