Speaker
Description
Fluorescence lifetime imaging microscopy (FLIM) is a powerful technique used to probe the local environment of fluorophores. Phasor analysis is a fit-free technique based on a FFT transformation of the intensity decay that provides a visual distribution of the molecular species, clustering pixels with similar lifetimes even when they are spatially separated in the image. Phasor analysis is increasingly being used due to its ease of interpretation. Here, we present Fluorescence Lifetime Ultimate Explorer (FLUTE), an open-source graphical user interface (GUI) for phasor analysis of FLIM data programmed in Python [1,2]. FLUTE simplifies and automates many aspects of the analysis of FLIM data acquired in the time domain, such as calibrating the FLIM data, performing interactive exploration of the phasor plot, displaying phasor plots and FLIM images with different lifetime contrasts simultaneously, and calculating the distance from known molecular species. After applying desired filters and thresholds, the final edited datasets can be exported for further user-specific analysis. FLUTE has been tested using several FLIM datasets including autofluorescence of zebrafish embryos and in vitro cells. 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.
References:
[1] D Gottlieb, B Asadipour, P. Kostina, TPL Ung, C Stringari. FLUTE: a Python GUI for interactive phasor analysis of FLIM data. Gottlieb D, Asadipour B, Kostina P, Ung TPL, Stringari C. FLUTE: A Python GUI for interactive phasor analysis of FLIM data. Biological Imaging. 2023;3:e21.
[2] https://github.com/LaboratoryOpticsBiosciences/FLUTE
| Authors | Dale Gottlieb , Bahar Asadipour , Polina Kostina , Thi Phuong Lien Ung and Chiara Stringari* |
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| Keywords | Data visualization, fluorescence lifetime microscopy, FLIM, phasor analysis, Python |
