Significance: Fluorescence lifetime imaging microscopy (FLIM) measures the decay rate of fluorophores, thus providing insights into molecular interactions. FLIM is a powerful molecular imaging technique that is widely used in biology and medicine. Aim: This perspective highlights some of the major advances in FLIM instrumentation, analysis, and biological and clinical applications that we have found impactful over the last year. Approach: Innovations in FLIM instrumentation resulted in faster acquisition speeds, rapid imaging over large fields of view, and integration with complementary modalities such as single-molecule microscopy or light-sheet microscopy. There were significant developments in FLIM analysis with machine learning approaches to enhance processing speeds, fit-free techniques to analyze images without a priori knowledge, and open-source analysis resources. The advantages and limitations of these recent instrumentation and analysis techniques are summarized. Finally, applications of FLIM in the last year include label-free imaging in biology, ophthalmology, and intraoperative imaging, FLIM of new fluorescent probes, and lifetime-based Förster resonance energy transfer measurements. Conclusions: A large number of high-quality publications over the last year signifies the growing interest in FLIM and ensures continued technological improvements and expanding applications in biomedical research.

中文翻译：

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用于生物学和医学的荧光寿命成像显微镜的最新创新

意义：荧光寿命成像显微镜 (FLIM) 可测量荧光团的衰减率，从而深入了解分子间的相互作用。FLIM 是一种强大的分子成像技术，广泛应用于生物学和医学。目标：这一观点突出了我们在过去一年中发现有影响的 FLIM 仪器、分析以及生物学和临床应用方面的一些重大进展。方法：FLIM 仪器的创新带来了更快的采集速度、大视场的快速成像以及与单分子显微镜或光片显微镜等互补模式的集成。FLIM 分析有了重大发展，使用机器学习方法来提高处理速度，在没有先验知识的情况下分析图像的无拟合技术，和开源分析资源。总结了这些最新的仪器和分析技术的优点和局限性。最后，去年 FLIM 的应用包括生物学、眼科和术中成像中的无标记成像、新型荧光探针的 FLIM 以及基于寿命的 Förster 共振能量转移测量。结论：去年大量高质量的出版物表明人们对 FLIM 的兴趣日益浓厚，并确保了技术的持续改进和生物医学研究中应用的扩大。新荧光探针的 FLIM，以及基于寿命的 Förster 共振能量转移测量。结论：去年大量高质量的出版物表明人们对 FLIM 的兴趣日益浓厚，并确保了技术的持续改进和生物医学研究中应用的扩大。新荧光探针的 FLIM，以及基于寿命的 Förster 共振能量转移测量。结论：去年大量高质量的出版物表明人们对 FLIM 的兴趣日益浓厚，并确保了技术的持续改进和生物医学研究中应用的扩大。