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。

Because of the high sensitivity and specificity and unparalleled ability to quantitatively analyze tissue microenvironments, fluorescence lifetime imaging microscopy (FLIM) is widely used in the field of life sciences. However, the use of FLIM in live imaging is hindered by its slow imaging speed. Recent advancements in optoelectronics and artificial intelligence technologies have ushered in a new era for in vivo FLIM. This review introduces ways to improve the imaging speed of the time-domain and frequency-domain FLIM techniques through hardware optimization and algorithmic improvements as well as via advancements in fundamental biomedical research and clinical diagnostics. Finally, we explore the future directions of the in vivo FLIM technology.