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High-throughput multiplexed fluorescence lifetime microscopy

Zhimin Zhang, Xin Liu, Minfei He, Yuran Huang, Liang Xu, Yubing Han, Dazhao Zhu, Xiang Hao, Cuifang Kuang, and Xu Liu

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Abstract

Fluorescence lifetime microscopy has been widely used in quantifying cellular interaction or histopathological identification of different stained tissues. A novel, to the best of our knowledge, approach for high-throughput multiplexed fluorescence lifetime imaging is presented. To establish a high-throughput fluorescence lifetime acquisition system, a uniformed illumination optical focus array was generated by a novel computer-generated hologram algorithm based on matrix triple product. This, in conjunction with an array detector and multichannel time-correlated single-photon counting, enables the full use of the acquisition ability of each detector. By utilizing interval segmentation of photon time detection, a high-throughput multiplexed fluorescence lifetime imaging is achieved. Experimental results demonstrate that this method achieves a fivefold increase in the collection throughput of fluorescence lifetime and is capable of simultaneous dual-target fluorescence lifetime measurement.

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Data underlying the results presented in this paper are not publicly available at this time but may be obtained from the authors upon reasonable request.

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