Fluorescence Lifetime Imaging (FLIM) in life sciences based on ultrashort laser scanning microscopy and time-correlated single photon counting (TCSPC) started 30 years ago in Jena/East-Germany. One decade later, first two-photon FLIM images of a human finger were taken with a lab microscope based on a tunable femtosecond Ti:sapphire laser. In 2002/2003, first clinical non-invasive two-photon FLIM studies on patients with dermatological disorders were performed using a novel multiphoton tomograph. Current in vivo two-photon FLIM studies on human subjects are based on TCSPC and focus on (i) patients with skin inflammation and skin cancer as well as brain tumors, (ii) cosmetic research on volunteers to evaluate anti-ageing cremes, (iii) pharmaceutical research on volunteers to gain information on in situ pharmacokinetics, and (iv) space medicine to study non-invasively skin modifications on astronauts during long-term space flights. Two-photon FLIM studies on volunteers and patients are performed with multiphoton FLIM tomographs using near infrared femtosecond laser technology that provide rapid non-invasive and label-free intratissue autofluorescence biopsies with picosecond temporal resolution.

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评论：临床体内多光子FLIM层析成像。

基于超短激光扫描显微镜和与时间相关的单光子计数（TCSPC）的生命科学中的荧光寿命成像（FLIM）于30年前在德国耶拿（Jena / East-Germany）开始。十年后，基于可调飞秒Ti：蓝宝石激光器的实验室显微镜拍摄了人手指的前两个光子FLIM图像。在2002/2003年，首次使用新型多光子断层扫描仪对皮肤病患者进行了临床非侵入性双光子FLIM研究。当前针对人类受试者的体内双光子FLIM研究基于TCSPC，并专注于（i）患有皮肤炎症和皮肤癌以及脑瘤的患者，（ii）对志愿者进行美容研究以评估抗衰老奶油的研究，（iii ）针对志愿者的药物研究，以获取有关原位药代动力学的信息，（iv）太空医学，以研究长期太空飞行中宇航员对皮肤的无创治疗。使用近红外飞秒激光技术的多光子FLIM断层扫描仪对志愿者和患者进行双光子FLIM研究，该技术可提供皮秒时间分辨率的快速无创且无标签的组织内自体荧光活检。