21 February 2017 Simultaneous acquisition of trajectory and fluorescence lifetime of moving single particles
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Abstract
Fluorescence lifetime imaging (FLIM) has been a powerful tool in life science because it can reveal the interactions of an excited fluorescent molecule and its environment. The combination with two-photon excitation (TPE) and timecorrelated single photon counting (TCSPC) provides it the ability of optical sectioning, high time resolution and detection efficiency. In previous work, we have introduced a two-dimensional acousto-optic deflector (AOD) into TCSPC-based FLIM to achieve fast and flexible FLIM. In this work, we combined the AOD-FLIM system with a single particle tracking (SPT) setup and algorithm and developed an SPT-FLIM system. Using the system, we acquired the trajectory and fluorescence lifetime of a moving particle simultaneously and reconstructed a life-time-marked pseudocolored trajectory, which might reflect dynamic interaction between the moving particle and its local environment along its motion trail. The results indicated the potential of the technique for studying the interaction between specific moving biological macromolecules and the ambient micro-environment in live cells.
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Qianqian Wu, Qianqian Wu, Jing Qi, Jing Qi, Danying Lin, Danying Lin, Wei Yan, Wei Yan, Rui Hu, Rui Hu, Xiao Peng, Xiao Peng, Junle Qu, Junle Qu, } "Simultaneous acquisition of trajectory and fluorescence lifetime of moving single particles", Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 1006922 (21 February 2017); doi: 10.1117/12.2251412; https://doi.org/10.1117/12.2251412
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