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Precision Fluorescence-Lifetime Imaging of a Moving Object

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We demonstrate precision lifetime analysis on FLIM data of a moving object. The technique is based on temporal-mosaic recording and image segmentation by the phasor plot of bh SPCImage NG data analysis software. A cluster of phasors is selected in the phasor space, identifying pixels of a given decay signature in the FLIM mosaic. These pixels are back-annotated in the mosaic, selecting parts of the objects irrespectively of their location in the individual images. The decay data of the pixels within the selected areas are summed up. The result is a single decay curve with extremely high pixel number which can be analysed at high precision.

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