Highly multiplexed spectral FLIM via physics informed data analysis.

ABSTRACT: Spectral fluorescence lifetime imaging (S-FLIM) allows for the simultaneous deconvolution of signal from multiple fluorophore species by leveraging both spectral and lifetime information. However, existing analyses still face multiple difficulties in decoding information collected from typical S-FLIM experiments. These include: using information from pre-calibrated spectra in environments that may differ from the cellular context in which S-FLIM experiments are performed; limitations in the ability to deconvolute species due to overlapping spectra; high photon budget requirements, typically about a hundred photons per pixel per species. Yet information on the spectra themselves are already encoded in the data and do not require pre-calibration. What is more, efficient photon-by-photon anal

SUBMITTER: Fazel M 

PROVIDER: S-EPMC12363814 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature