Time‐correlated single photon counting is the “gold‐standard” method for fluorescence lifetime measurements and has demonstrated potential for clinical deployment. However, the translation of the technology into clinic is hindered by the use of ultrasensitive detectors, which make the fluorescence acquisition impractical with bright lighting conditions such as in clinical settings. We address this limitation by interleaving periodic fluorescence detection with synchronous out‐of‐phase externally modulated light source, thus guaranteeing specimen illumination and a fluorescence signal free from bright background light upon temporal separation. Fluorescence lifetime maps are generated in real‐time from single‐point measurements by tracking a reference beam and using the phasor approach. We demonstrate the feasibility and practicality of this technique in a number of biological specimens, including real‐time mapping of degraded articular cartilage. This method is compatible and can be integrated with existing clinical microscopic, endoscopic and robotic modalities, thus offering a new pathway towards label‐free diagnostics and surgical guidance in a number of clinical applications.

中文翻译：

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具有同步外部照明的基于光纤的实时荧光寿命成像：临床翻译的新途径。

与时间相关的单光子计数是荧光寿命测量的“金标准”方法，已证明具有临床应用潜力。但是，使用超灵敏检测器阻碍了该技术向临床的转化，这使荧光采集在明亮的照明条件下（例如在临床环境中）变得不切实际。我们通过将周期性荧光检测与同步异相外部调制光源进行交错处理来解决这一局限性，从而确保样品照明和荧光信号在暂时分离时没有明亮的背景光。通过跟踪参考光束并使用相量方法，从单点测量实时生成荧光寿命图。我们在许多生物学标本中证明了该技术的可行性和实用性，包括对退化的关节软骨的实时定位。该方法是兼容的，并且可以与现有的临床显微镜，内窥镜和机器人模式相集成，从而为许多临床应用中的无标签诊断和手术指导提供了一条新途径。