Abstract
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.