FLIM Playground: An interactive, end-to-end graphical user interface for analyzing single cells with fluorescence lifetime imaging microscopy
Year: 2025
Type: preprint
Abstract: Fluorescence lifetime imaging microscopy (FLIM) is a cellular-resolution molecular imaging technique. Yet, the journey from raw photon decays to biological insight remains fragmented by multi-step data extraction and siloed analyses. This work presents FLIM Playground, the first interactive graphical platform that unifies single-cell FLIM workflows, embeds user checks at each stage, and offers diverse user options. Built in Python and available open-source, FLIM Playground runs on major operating systems as a ready-to-run application and is web deployable. Its Data Extraction section collects and checks field-of-view metadata, calibrates via instrument response function shift or fluorescence lifetime standard, and extracts single-cell fluorescence lifetime features, along with morphology and texture features across channels. Multiple datasets can be merged through an interface that assigns categorical labels. The Data Analysis section provides real-time visual analytic modules for outputs from Data Extraction or user-provided datasets. Lifetime extraction by fitting and phasor were validated by comparison with a commercial software and published results, respectively, and both sections were demonstrated on a FLIM dataset of cancer cell lines to obtain biological insights. By adopting best practices and offering interactivity, FLIM Playground accelerates hypothesis-driven discovery and promotes reproducibility, and its modular design can incorporate new imaging modalities, extraction methods, and analysis modules.
Authors: Wenxuan Zhao, Kayvan Samimi, Melissa C. Skala, Rupsa Datta
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: green
Awards: Collaborative Research: EAGER: Autofluorescence lifetime flow sorting with time-correlated single photon counting, G1923404860, G2692556688, Optical imaging to predict cell-level genetic heterogeneity and treatment sensitivity in colorectal cancer, G6059030093, G7096540765, Label-free imaging of CAR T cell metabolism, Functional optical imaging for rapid, label-free predictions of treatment response and clonal evolution in patient-derived cancer organoids, Label-free single-cell imaging for quality control of cardiomyocyte biomanufacturing
OpenAlex ID: W4414764690