Department of Biomedical Engineering, University of California, Davis, CA 95616, USA.
Microsc Microanal. 2013 Aug;19(4):791-8. doi: 10.1017/S1431927613001530. Epub 2013 May 23.
A clinically compatible fluorescence lifetime imaging microscopy (FLIM) system was developed. The system was applied to intraoperative in vivo imaging of head and neck squamous cell carcinoma (HNSCC). The endoscopic FLIM prototype integrates a gated (down to 0.2 ns) intensifier imaging system and a fiber-bundle endoscope (0.5-mm-diameter, 10,000 fibers with a gradient index lens objective 0.5 NA, 4-mm field of view), which provides intraoperative access to the surgical field. Tissue autofluorescence was induced by a pulsed laser (337 nm, 700 ps pulse width) and collected in the 460 ± 25 nm spectral band. FLIM experiments were conducted at 26 anatomic sites in ten patients during head and neck cancer surgery. HNSCC exhibited a weaker florescence intensity (~50% less) when compared with healthy tissue and a shorter average lifetime (τ(HNSCC) = 1.21 ± 0.04 ns) than the surrounding normal tissue (τN = 1.49 ± 0.06 ns). This work demonstrates the potential of FLIM for label-free head and neck tumor demarcation during intraoperative surgical procedures.
开发了一种临床兼容的荧光寿命成像显微镜（FLIM）系统。该系统应用于头颈部鳞状细胞癌（HNSCC）的术中体内成像。内窥镜 FLIM 原型集成了门控（低至 0.2 ns）增强器成像系统和纤维束内窥镜（0.5 毫米直径，带有梯度折射率透镜物镜 0.5 NA 的 10,000 根光纤，4 毫米视野），为手术区域提供了术中通道。组织自发荧光由脉冲激光器（337nm，700ps 脉冲宽度）激发，并在 460±25nm 的光谱带中收集。在十名头颈部癌症手术患者的 26 个解剖部位进行了 FLIM 实验。与健康组织相比，HNSCC 的荧光强度较弱（约低 50%），平均寿命较短（τ（HNSCC）=1.21±0.04ns），而周围正常组织的平均寿命较长（τ N =1.49±0.06ns）。这项工作证明了 FLIM 对头颈部肿瘤在术中手术过程中进行无标记肿瘤边界区分的潜力。
