Structured illumination multispectral fluorescence lifetime microendoscopy to detect oral premalignancies

 描述(申请人提供)：美国癌症协会估计，美国每年报告的口腔和咽部浸润性癌症新病例超过4万例，晚期疾病的五年生存率仍低于40%。本研究的目的是设计、制造和测试一种用于口腔癌前病变活体检测的结构照明多光谱广域荧光寿命成像显微内窥镜。将结构照明(SI)与荧光寿命成像(FLIM)相结合，将能够生成高保真的三维体积的荧光强度和寿命 上皮组织。我们相信，由于结构蛋白和代谢辅助因子等分子标志物在组织中的不同分布，收集跨上皮细胞的光学切片的能力将允许表征组织的生化成分，并最终将有助于对早期口腔癌发展的高敏感性和特异性检测。该方案的具体目标是：1)构建提供光学深度切片的结构照明多光谱薄膜内窥镜系统；2)通过体外和体外成像来表征结构照明多光谱薄膜内窥镜的性能并优化其设计指标；3)通过体内组织成像来验证和演示结构照明多光谱薄膜内窥镜的性能。这项工作的成功完成将使第一个能够选择性地成像组织内特定深度的多光谱SI-FILM显微内窥镜的开发成为可能，并将展示其在口腔癌前病变和恶性病变的非侵入性表征方面的潜力。由于在多个光谱通道中收集的荧光强度和寿命中编码了丰富的特定分子信息，这种设备除了可以提供临床医生的主观评估外，还可以用于提供口腔的客观筛查。虽然所提出的内窥镜是为在口腔中应用该设备而设计的，但只要对内窥镜稍作修改，SI-FLIM系统也可以适用于检测和诊断其他癌症(如基底细胞癌、结直肠癌和子宫颈癌)。

 DESCRIPTION (provided by applicant): The American Cancer Society estimates that more than 40,000 new cases of invasive cancer in the oral cavity and pharynx are reported each year in the US, and five-year survival rates remain below 40% for advanced stage disease. The goal of this research is to design, build and test a structured illumination-enabled multispectral widefield fluorescence lifetime imaging microendoscope for detection of oral precancer in vivo. Combining structured illumination (SI) with fluorescence lifetime imaging (FLIM) would enable generation of high fidelity three dimensional volumes of the fluorescence intensity and lifetime of the epithelial tissue. We believe that the ability to collect 'optical sections' across the epithelum would allow characterization of tissue biochemical composition due to differential distribution of molecular markers, such as structural proteins and metabolic cofactors, within tissue and will ultimately aid in high sensitivity and specificity detection of early oral cancer development. The specific aims of this proposal are to: 1) construct a structured illumination multispectral FLIM endoscope (SI-FLIM) system to provide optical depth sectioning; 2) characterize the performance and optimize the design specifications of the SI-FLIM endoscope by imaging in vitro and ex vivo; and 3) validate and demonstrate the capability of SI-FLIM by imaging tissue in vivo. The successful completion of this work will enable the development of the first multispectral SI-FLIM microendoscope with the ability to selectively image specific depths within the tissue, and will demonstrate its potential for non-invasive characterization of oral premalignant and malignant lesions. With the wealth of specific molecular information encoded in the collected fluorescence intensity and lifetime in multiple spectral channels, such a device can serve to provide an objective screening of the oral cavity in addition to the clinician's subjective evaluation. Although the proposed endoscope is designed for application of this device in the oral cavity, the SI-FLIM system can be applicable to detect and diagnose other carcinomas (e.g. basal cell, colorectal, and cervical) as well with slight modifications to the endoscope.