FLIMExplorer: interactive GUI for object-based visualization and analysis of fluorescence lifetime images

Area of Science:

• Biophysics
• Cellular Metabolism
• Microscopy

Background:

• Fluorescence lifetime imaging microscopy (FLIM) offers non-invasive, high-resolution cellular metabolism analysis.
• Current FLIM workflows are complex, fragmented, and lack integration between quantitative data and image context, hindering quality control.

Purpose of the Study:

• To introduce FLIMExplorer, an interactive Python-based tool for streamlined single-cell FLIM data analysis and visualization.
• To bridge the gap between quantitative FLIM measurements and their corresponding image data, enhancing transparency and quality control.

Main Methods:

• FLIMExplorer processes pixel-level FLIM outputs or pre-processed cell-average datasets.
• It integrates quantitative FLIM endpoints (e.g., NAD(P)H and FAD lifetimes and amplitudes) with image visualization.
• The tool provides a graphical user interface for exploring single-cell features and performing statistical comparisons.

Main Results:

• FLIMExplorer links quantitative FLIM data points to their specific image objects.
• Users can visualize and analyze key FLIM features at the single-cell level within a unified platform.
• The tool facilitates statistical comparisons across experimental groups, improving data interpretation.

Conclusions:

• FLIMExplorer enhances downstream FLIM analysis by integrating visualization, quality control, and statistical analysis.
• It provides a more transparent and context-aware approach to single-cell metabolic profiling using FLIM.
• This tool improves the interpretability and reproducibility of FLIM-based cellular metabolism studies.