Endoscopic Fluorescence Lifetime Imaging for In Vivo Intraoperative Diagnosis of Oral Carcinoma

Abstract 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 (τHNSCC = 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.

[1]  Laura Marcu,et al.  Fluorescence Lifetime Techniques in Medical Applications , 2012, Annals of Biomedical Engineering.

[2]  Laura Marcu,et al.  Dynamic tissue analysis using time- and wavelength-resolved fluorescence spectroscopy for atherosclerosis diagnosis , 2011, Optics express.

[3]  K. Svanberg,et al.  Malignant tumor and atherosclerotic plaque diagnosis using laser-induced fluorescence , 1990 .

[4]  Frank Y. S. Chuang,et al.  Fluorescence lifetime imaging microscopy: in vivo application to diagnosis of oral carcinoma. , 2009, Optics letters.

[5]  N Ramanujam,et al.  In vivo fluorescence spectroscopy: potential for non-invasive, automated diagnosis of cervical intraepithelial neoplasia and use as a surrogate endpoint biomarker. , 1994, Journal of cellular biochemistry. Supplement.

[6]  J. Roodenburg,et al.  The status of in vivo autofluorescence spectroscopy and imaging for oral oncology. , 2005, Oral oncology.

[7]  Karthik Vishwanath,et al.  Fluorescence Spectroscopy In Vivo , 2011 .

[8]  E. Sevick-Muraca,et al.  Quantitative optical spectroscopy for tissue diagnosis. , 1996, Annual review of physical chemistry.

[9]  R. Cubeddu,et al.  Time-resolved fluorescence imaging in biology and medicine , 2002 .

[10]  Cristina Kurachi,et al.  Noninvasive evaluation of oral lesions using depth‐sensitive optical spectroscopy , 2009, Cancer.

[11]  Rebecca Richards-Kortum,et al.  Fluorescence spectroscopy of oral tissue: Monte Carlo modeling with site-specific tissue properties. , 2009, Journal of biomedical optics.

[12]  Qiyin Fang,et al.  Laguerre-based method for analysis of time-resolved fluorescence data: application to in-vivo characterization and diagnosis of atherosclerotic lesions. , 2006, Journal of biomedical optics.

[13]  J A Jo,et al.  Miniaturized side-viewing imaging probe for fluorescence lifetime imaging (FLIM): validation with fluorescence dyes, tissue structural proteins and tissue specimens , 2007, New journal of physics.

[14]  Qiyin Fang,et al.  Detection of rupture-prone atherosclerotic plaques by time-resolved laser-induced fluorescence spectroscopy. , 2009, Atherosclerosis.

[15]  Laura Marcu,et al.  A novel method for fast and robust estimation of fluorescence decay dynamics using constrained least-squares deconvolution with Laguerre expansion , 2017, Physics in medicine and biology.

[16]  Jan Siegel,et al.  Time-domain fluorescence lifetime imaging applied to biological tissue , 2004, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[17]  N. Ramanujam,et al.  In vivo multiphoton microscopy of NADH and FAD redox states, fluorescence lifetimes, and cellular morphology in precancerous epithelia , 2007, Proceedings of the National Academy of Sciences.

[18]  Laura Marcu,et al.  Multimodal characterization of compositional, structural and functional features of human atherosclerotic plaques , 2011, Biomedical optics express.

[19]  S Andersson-Engels,et al.  Multispectral tissue characterization with time-resolved detection of diffusely scattered white light. , 1993, Optics letters.

[20]  Laura Marcu,et al.  Fluorescence lifetime imaging for the characterization of the biochemical composition of atherosclerotic plaques. , 2011, Journal of biomedical optics.

[21]  Laura Marcu,et al.  In vivo validation of a bimodal technique combining time-resolved fluorescence spectroscopy and ultrasonic backscatter microscopy for diagnosis of oral carcinoma , 2012, Journal of biomedical optics.

[22]  J. Lakowicz.  Principles of fluorescence spectroscopy , 1983 .