Laser & Optoelectronics Progress, Volume. 62, Issue 18, 1817023(2025)

Free-Addressing FRET Technique Based on Fluorescent Lifetime Detection (Invited)

Jiahua Chen1,3,4, Huimin Jiang2,3,4, Yao Lu1,3,4, Kai Wen1,3,4, Sha An1,3,4, Peng Gao1,3,4, Xiaofang Wang1,3,4, Tanping Li1,3,4, Juanjuan Zheng1,3,4, Hongfei Suo1,3,4、*, Lixin Liu2,3,4, and Peng Gao1,3,4、**
Author Affiliations
  • 1School of Physics, Xidian University, Xi'an 710071, Shaanxi , China
  • 2School of Optoelectronic Engineering, Xidian University, Xi'an 710071, Shaanxi , China
  • 3Key Laboratory of Optoelectronic Perception of Complex Environment, Ministry of Education, Xi'an 710071, Shaanxi , China
  • 4Engineering Research Center of Information Nanomaterials, Universities of Shaanxi Province, Xi'an 710071, Shaanxi , China
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    Fluorescence resonance energy transfer (FRET) serves as a versatile technique for quantifying intermolecular interactions by measuring the energy transfer between donor and acceptor molecules. In this work, a free-addressing FRET technique based on fluorescence lifetime measurements is proposed. Within the framework of a dual-color confocal microscopy, this method utilizes two galvanometric scanners to freely address a focused light to any locations on a sample and uses a time-correlated single photon counting board to capture the trigger signals from the scanners, and the arrival times of photons relative to previous laser pulses. Consequently, the fluorescence lifetime at any region of interest within the field-of-view can be obtained. Once the fluorescent lifetimes of donor fluorophores before and after bleaching the acceptor fluorophores are determined, the FRET efficiency at the targeted site can be further calculated. Compared with the traditional intensity-based FRET measurement method, this approach does not require special calibration operations and significantly reduces measurement times from minutes to milliseconds. We can envisage that the proposed technique can be widely applied to life science, medical diagnosis, materials science, and so on.

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    Jiahua Chen, Huimin Jiang, Yao Lu, Kai Wen, Sha An, Peng Gao, Xiaofang Wang, Tanping Li, Juanjuan Zheng, Hongfei Suo, Lixin Liu, Peng Gao. Free-Addressing FRET Technique Based on Fluorescent Lifetime Detection (Invited)[J]. Laser & Optoelectronics Progress, 2025, 62(18): 1817023

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    Paper Information

    Category: Medical Optics and Biotechnology

    Received: Jul. 14, 2025

    Accepted: Aug. 4, 2025

    Published Online: Sep. 16, 2025

    The Author Email: Hongfei Suo (hongfeisuo@stu.xidian.edu.cn), Peng Gao (peng.gao@xidian.edu.cn)

    DOI:10.3788/LOP251659

    CSTR:32186.14.LOP251659

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