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FLIM Playground: An interactive, end-to-end graphical user interface for analyzing single cells with fluorescence lifetime imaging microscopy.

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Title: FLIM Playground: An interactive, end-to-end graphical user interface for analyzing single cells with fluorescence lifetime imaging microscopy.
Authors: Zhao W; Morgridge Institute for Research, Madison, WI, USA., Samimi K; Morgridge Institute for Research, Madison, WI, USA., Skala MC; Morgridge Institute for Research, Madison, WI, USA; Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA., Datta R; Morgridge Institute for Research, Madison, WI, USA. Electronic address: rdatta@morgridge.org.
Source: Cell reports methods [Cell Rep Methods] 2026 Aug 17; Vol. 6 (8), pp. 101484. Date of Electronic Publication: 2026 Jun 10.
Publication Type: Journal Article
Language: English
Journal Info: Publisher: Elsevier Inc Country of Publication: United States NLM ID: 9918227360606676 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2667-2375 (Electronic) Linking ISSN: 26672375 NLM ISO Abbreviation: Cell Rep Methods Subsets: MEDLINE
Imprint Name(s): Original Publication: [New York] : Elsevier Inc., [2021]-
MeSH Terms: Single-Cell Analysis*/methods , User-Computer Interface*, Microscopy, Fluorescence/methods ; Image Processing, Computer-Assisted/methods ; Humans ; Software ; Cell Line, Tumor
Abstract: Fluorescence lifetime imaging microscopy (FLIM) is sensitive to molecular environments and enables high-resolution mapping of cellular heterogeneity. Yet, the journey from raw photon decays to biological insight remains fragmented by multi-step data extraction and siloed analyses, creating burdens for experts and non-experts alike. This work presents FLIM Playground, the first interactive graphical platform that unifies single-cell FLIM workflows. Modularly designed to encompass data extraction (if desired) and data analysis, FLIM Playground can check field-of-view metadata, calibrate and extract fluorescence lifetime features per region of interest along with morphology and texture features across channels, merge multiple datasets, and provide real-time visual analytic modules. Lifetime extraction was validated against commercial software and published results, and both data extraction and analysis were demonstrated on a FLIM dataset of cancer cell lines. By adopting best practices and offering interactivity, FLIM Playground promotes reproducibility, allows for expansion to new imaging modalities, and accelerates hypothesis-driven discovery.
(Copyright © 2026 The Authors. Published by Elsevier Inc. All rights reserved.)
Competing Interests: Declaration of interests M.C.S. is an adviser to Elephas Biosciences. All other authors declare they have no competing interests.
Comments: Update of: bioRxiv. 2025 Nov 17:2025.09.30.679625. doi: 10.1101/2025.09.30.679625.. (PMID: 41256698)
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Grant Information: R01 CA272855 United States CA NCI NIH HHS; R01 CA278051 United States CA NCI NIH HHS; R01 HL165726 United States HL NHLBI NIH HHS; R37 CA226526 United States CA NCI NIH HHS
Contributed Indexing: Keywords: CP: imaging; FLIM; GUI; TCSPC; fluorescence lifetime imaging microscopy; image analysis; interactive visualization; lifetime fitting; metabolic imaging; open-source software; phasor analysis
Entry Date(s): Date Created: 20260610 Date Completed: 20260817 Latest Revision: 20260824
Update Code: 20260824
PubMed Central ID: PMC13494553
DOI: 10.1016/j.crmeth.2026.101484
PMID: 42269617
Database: MEDLINE
Description
ISSN:2667-2375
DOI:10.1016/j.crmeth.2026.101484