荧光寿命显微成像（FLIM）已经广泛应用于生命科学研究领域，具有高灵敏和高特异性的特点，在对组织微环境进行定量表征方面具有独特优势，但由于成像速度相对较慢，限制了FLIM的活体应用。近年来，随着光电子器件和人工智能等技术的发展，开启了FLIM活体成像新篇章。介绍通过优化硬件和算法两方面提升时域和频域FLIM技术的成像速度，以及其在生物医学基础研究和临床疾病诊断中的应用研究进展。最后，对活体FLIM成像的未来发展进行展望。

Fluorescence imaging in the second near-infrared window (NIR-II, 900–1880nm) with less scattering background in biological tissues has been combined with the confocal microscopic system for achieving deep in vivo imaging with high spatial resolution. However, the traditional NIR-II fluorescence confocal microscope with separate excitation focus and detection pinhole makes it possess low confocal efficiency, as well as difficultly to adjust. Two types of upgraded NIR-II fluorescence confocal microscopes, sharing the same pinhole by excitation and emission focus, leading to higher confocal efficiency, are built in this work. One type is fiber-pinhole-based confocal microscope applicable to CW laser excitation. It is constructed for fluorescence intensity imaging with large depth, high stabilization and low cost, which could replace multiphoton fluorescence microscopy in some applications (e.g., cerebrovascular and hepatocellular imaging). The other type is air-pinhole-based confocal microscope applicable to femtosecond (fs) laser excitation. It can be employed not only for NIR-II fluorescence intensity imaging, but also for multi-channel fluorescence lifetime imaging to recognize different structures with similar fluorescence spectrum. Moreover, it can be facilely combined with multiphoton fluorescence microscopy. A single fs pulsed laser is utilized to achieve up-conversion (visible multiphoton fluorescence) and down-conversion (NIR-II one-photon fluorescence) excitation simultaneously, extending imaging spectral channels, and thus facilitates multi-structure and multi-functional observation.

自2001年被发明以来，超导纳米线单光子探测器（SNSPD）迅速成长为近红外波段的明星光子探测器，其在近红外波段如1550 nm处系统探测效率超过95%，暗计数率低于1 cps（counts per second），时间抖动优于10 ps，探测速率高于1 GHz，并广泛应用在量子信息领域。近年来，研究人员开始将SNSPD引入到生物领域，以替代在近红外波段具有低信噪比、多后脉冲的半导体单光子探测器。本文将介绍SNSPD的探测原理和性能指标，并系统地阐述SNSPD在生物领域中的应用现状和发展前景。

Apoptosis is very important for the maintenance of cellular homeostasis and is closely related to the occurrence and treatment of many diseases. Mitochondria in cells play a crucial role in programmed cell death and redox processes. Nicotinamide adenine dinucleotide (NAD(P)H) is the primary producer of energy in mitochondria, changing NAD(P)H can directly reflect the physiological state of mitochondria. Therefore, NAD(P)H can be used to evaluate metabolic response. In this paper, we propose a noninvasive detection method that uses two-photon fluorescence lifetime imaging microscopy (TP-FLIM) to characterize apoptosis by observing the binding kinetics of cellular endogenous NAD(P)H. The result shows that the average fluorescence lifetime of NAD(P)H and the fluorescence lifetime of protein-bound NAD(P)H will be affected by the changing pH, serum content, and oxygen concentration in the cell culture environment, and by the treatment with reagents such as H2O2 and paclitaxel. Taxol (PTX). This noninvasive detection method realized the dynamic detection of cellular endogenous substances and the assessment of apoptosis.

有机-无机卤化铅钙钛矿多晶薄膜太阳能电池在近几年的研究中实现了光电转换效率的快速增长。然而，其多晶结构的活性层导致器件仍然遭受到表面和晶界位置缺陷引起的性能衰减。本研究借助两种有机氢碘酸盐，即苯乙基碘化胺(Phenethylammonium iodide,PEAI)和邻氟苯乙胺碘(2-Fluorophenylethylammonium iodide,o-F-PEAI)，在CH3NH3PbI3钙钛矿多晶薄膜表面形成钝化层。扫描电子显微镜和原子力显微镜分析结果显示，PEAI和o-F-PEAI处理后的钙钛矿薄膜晶界被钝化层明显填充，表面粗糙度也显著下降。另外，荧光寿命成像分析结果显示钝化后的钙钛矿薄膜具有更多的光子数和更长的荧光寿命。上述结果表明，PEAI和o-F-PEAI诱导的钝化层可以有效抑制多晶薄膜表面和晶界位置的缺陷复合行为。因此，钝化后的倒置结构钙钛矿太阳能电池器件功率转换效率(Power conversion efficiency,PCE)可以达到21%。此外，o-F-PEAI钝化处理后的器件由于氟离子的作用表现出更好的器件稳定性。

细胞是动植物结构和生命活动的基本单位。 细胞过程的一个重要特点就是其生化组分在时空调控上的相互作用关系。 然而, 利用传统的生化方法（如酵母双杂交系统、 pull-down系统等）很难在空间上评估活细胞内分子间的相互作用。 光学技术的快速发展, 为研究活细胞中生物分子的时空动态提供了新的遗传研究工具, 其中荧光共振能量转移-荧光寿命显微成像（FRET-FLIM）技术在实时探测分析活细胞中生物大分子构象变化和分子间动态相互作用过程具有独特的优势, 如: 实现对活细胞的实时“可视化”研究, 同时具有高时空分辨率; 检测更加灵敏、 结果可信度高; 且基于简易的数学运算完成简单快捷的分析程序。 介绍FRET-FLIM技术的理论背景知识, 对比了该技术与传统蛋白相互作用技术研究的利弊, 同时归纳了其在蛋白相互作用、 细胞生物学和疾病诊断等方面的最新应用研究进展, 最后总结和讨论了FRET-FLIM技术的未来发展趋势, 以期能够为揭示活细胞的结构和细胞过程相关研究提供新的见解。