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Updated: Nov 14, 2025

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Published on: February 9, 2012
Theoretical investigations of a modified compressed ultrafast photography method suitable for single-shot
Abstract:
A single-shot fluorescence lifetime imaging (FLIM) method based on the compressed ultrafast photography (CUP) is proposed, named space-restricted CUP (srCUP). srCUP is suitable for imaging objects moving slowly (<∼150/Mmm/s, M is the magnification of the objective lens) in the field of view with the intensity changing within nanoseconds in a measurement window around 10 ns. We used synthetic datasets to explore the performances of srCUP compared with CUP and TCUP (a variant of CUP). srCUP not only provides superior reconstruction performances, but its reconstruction speed is also twofold and threefold faster than CUP and TCUP, respectively. The lifetime determination performances were assessed by estimating lifetime components, amplitude- and intensity-weighted average lifetimes (τA and τI), with the reconstructed scenes using the least squares method based on a bi-exponential model. srCUP has the best accuracy and precision for lifetime determinations with a relative bias less than 7% and a coefficient of variation less than 7% for τA, and a relative bias less than 10% and a coefficient of variation less than 11% for τI.
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