FLUTE: a Python GUI for interactive phasor analysis of FLIM data
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Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.03.31.534529v1?rss=1

Authors: Gottlieb, D., Asadipour, B., Ung, T. P. L., Stringari, C.

Abstract: Fluorescence lifetime imaging microscopy (FLIM) is a powerful technique used to probe the local environment of fluorophores. The phasor approach to FLIM data is a fit-free analysis and is increasingly used due to its ease of interpretation. To date, no open-source graphical user interface (GUI) for phasor analysis of FLIM data is available thus limiting the widespread use of phasor analysis in biomedical research. Here we present (F)luorescence (L)ifetime (U)l(t)imate (E)xplorer (FLUTE), a Python GUI that is designed to fill this gap. FLUTE simplifies and automates many aspects of FLIM analysis, such as calibrating the FLIM data, performing interactive exploration of the phasor plot with cursors, displaying the phasor plot and the FLIM images with different lifetime contrasts and calculating the relative concentration of molecular species. The final edited data sets after applying the desired filters and thresholds can be exported for further user specific analysis. FLUTE was tested using several FLIM data sets including autofluorescence of Zebrafish embryos, cells in vitro and intact live tissues. In summary our user-friendly GUI extends the advantages of phasor plotting by making the data visualization and analysis easy and interactive, allows for analysis of large FLIM datasets and accelerates FLIM analysis for non-specialized labs.

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