High-throughput multiplexed fluorescence lifetime microscopy

• DOI: 10.1364/ol.503136
• Publication Date: 2023-09-29T13:00:15Z
• Authors: Zhimin Zhang; Xin Liu; Minfei He; Yuran Huang; Liang Xu; Yubing Han; Dazhao Zhu; Xiang Hao; Cuifang Kuang; Xu Liu

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.