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13 March 2024 Fast analysis with minimal user interaction in fluorescence lifetime imaging
Felix Koberling, Fabio Barachati, Marcelle König, Maria Loidolt-Krueger, Ellen Schmeyer, Matthias Patting, Marcus Sackrow, Uwe Ortmann, Evangelos Sisamakis, Rainer Erdmann
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Proceedings Volume PC12849, Single Molecule Spectroscopy and Superresolution Imaging XVII; PC128490N (2024) https://doi.org/10.1117/12.3001448
Event: SPIE BiOS, 2024, San Francisco, California, United States
Abstract
Fluorescence Lifetime Imaging (FLIM) has become more attractive in recent years as it offers increased specificity in many assays as well as the possibility of multiplexing the read out of many markers with a small number of detectors. Here we present how FLIM modalities are implemented in Luminosa, the new single-photon counting confocal microscope by PicoQuant. Thanks to a dynamic binning format and GPU-based algorithms FLIM images of 1024x1024 can be analysed in a few seconds. The FLIM analysis workflow suggests the best fitting model based on statistical arguments and requires minimal user interaction making these modalities become accessible to new users who can then confidently start working with FLIM and incorporate it into their research toolbox combining the strengths of phasor plots with decay fitting.
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Felix Koberling, Fabio Barachati, Marcelle König, Maria Loidolt-Krueger, Ellen Schmeyer, Matthias Patting, Marcus Sackrow, Uwe Ortmann, Evangelos Sisamakis, and Rainer Erdmann "Fast analysis with minimal user interaction in fluorescence lifetime imaging", Proc. SPIE PC12849, Single Molecule Spectroscopy and Superresolution Imaging XVII, PC128490N (13 March 2024); https://doi.org/10.1117/12.3001448
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KEYWORDS
Fluorescence lifetime imaging
Analytical research
Statistical analysis
Confocal microscopy
Detection and tracking algorithms
Microscopes
Multiplexing
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