Nonlinear Optical Endomicroscopy for Optical Biopsy of Cancer in Internal Organs

用于内脏器官癌症光学活检的非线性光学内镜检查

• 批准号: 8213488
• 负责人: Xingde Li
• 金额: $ 55.67万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-20 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8213488
• 关键词: Academia; Address; Animal Model; Area; Biochemical; Biological; Biomedical Engineering; Biopsy; Caliber; Cancer Detection; Clinic; Clinical; Collection; Computers; Detection; Development; Diagnosis; Diagnostic; Endoscopes; Engineering; Esophagus; Excision; Family suidae; Fiber; Fiber Optics; Financial compensation; Florida; Fluorescence; Fluorescent Dyes; Gastroenterology; Gastrointestinal tract structure; Gastroscopes; Generations; Histology; Histopathology; Human; Image; Image Guided Biopsy; Imagery; Imaging technology; In Situ; Industry; Intervention; Laboratories; Lasers; Length; Life; Malignant Neoplasms; Medicine; Metabolic; Methods; Microscope; Microscopy; Mucous Membrane; NADH; Neoplasms; Operative Surgical Procedures; Optical Biopsy; Optics; Organ; Pathology; Performance; Physiologic pulse; Play; Probability; Research; Research Personnel; Resolution; Scanning; Screening for cancer; Sensitivity and Specificity; Signal Transduction; Silicon Dioxide; Specimen; Speed; Technology; Testing; Therapeutic; Time; Tissues; Translating; Treatment outcome; Universities; Work; angiogenesis; base; cancer imaging; clinical application; clinical practice; engineering design; flexibility; human tissue; improved; in vivo; interest; lens; multidisciplinary; novel; novel strategies; operation; optical fiber; optical imaging; programs; public health relevance; second harmonic; tool; tumor; two-photon; voltage

DESCRIPTION (provided by applicant): There is a critical clinical need for a noninvasive high-resolution imaging technology for early cancer detection and guidance of biopsy in internal organs. Two-photon fluorescence (TPF) and second harmonic generation (SHG) microscopy is a powerful technology to address the above clinical need by providing structural and biochemical/metabolic information about biological tissues at subcellular resolution without the need for tissue removal or external fluorescent agents. However, its in vivo clinical application remains extremely limited due to the lack of a miniature technology platform. The objective of this multidisciplinary proposal is to develop an all-fiber-optic scanning endomicroscopy technology which is able to bring TPF/SHG microscopy to clinic for internal organ imaging. It involves 5 partners with 2 from academia and 3 from industry. The proposed technology will integrate all essential functions of a scanning laser microscope into a single flexible fiber-optic probe of a small diameter (~2.4-3.4 mm), with built-in mechanisms for femtosecond pulse delivery, dispersion management, nonlinear effect suppression, beam focusing, rapid 2D raster beam scanning, TPF/SHG collection, and focus tracking (or depth scanning). The small size permits its integration with a standard red-flagging technology (such as a gastroscope). In this proposal, we plan to tackle the major challenges in developing such an endomicroscopy technology and evaluate its feasibility for subcellular resolution imaging and for cancer detection. The Specific Aims are to: (1) Develop new double-clad fibers (DCF) of a pure silica core, large inner clad and numerical aperture to dramatically suppress the in-fiber TPF/SHG background (e.g. by 50 folds) and improve TPF/SHG collection efficiency (e.g. by ~15 folds) over commercially available DCFs; (2) Develop a super-achromatic microlens of a 2.1mm diameter and 0.6 NA to improve the TPF/SHG collection efficiency by at least 20 folds over a GRIN lens; (3) Explore a novel approach based on high-order mode DCFs to suppress nonlinear effects in optic fiber and improve TPF/SHG excitation probability; (4) Develop novel MEMS scanners of a small footprint (1.6 x 1.6 mm) and an extremely low drive voltage (10V max) to achieve rapid 2D raster beam scanning and real-time focus tracking. A fully integrated endomicroscope with customized DCFs, microlens and MEMS scanners will be developed, capable of 3D TPF/SHG imaging; (5) Conduct in vivo endoscopic TPF/SHG imaging of swine esophagus to evaluate the performance, and design, engineering and operation issues of the scanning probe; and (6) Evaluate the feasibility of the proposed technology for cancer detection and tumor margin identification using ex vivo human esophagus specimens, and correlate TPF/SHG endomicroscopy images with corresponding histology. The significance of the proposed research is to translate the powerful TPF/SHG microscopy technology to clinical practice for cancer detection and image-guided biopsy in internal organs, and enable noninvasive real-time visualization of tissue histopathology in situ to significantly improve diagnostic and biopsy yields. In addition, the proposed endomicroscopy technology will also be applicable (although outside the scope of this proposal) to many other clinical scenarios such as for guidance of surgical interventions and for in vivo assessment of metabolic function of living tissues. PUBLIC HEALTH RELEVANCE: The objective of this multidisciplinary proposal is to develop and test an all-fiber-optic 3D scanning endomicroscopy technology, which can bring the powerful bench-top TPF/SHG microscopy technology to clinical practice for imaging internal organs (such as the gastrointestinal tract) that was not previously possible with TPF/SHG microscopy. The proposed technology can function in a form of "optical biopsy" by providing structural and biochemical/metabolic information about biological tissues at subcellular resolution without the need for tissue removal or external fluorescent agents. Successful completion of the proposed research will bring us a new tool, which enables visualization of histopathology in situ and improves our capability for early cancer detection, tumor margin identification, and guidance of biopsy and interventions.

描述（由申请人提供）：对于早期癌症检测和内部器官活检的指导，对非侵入性高分辨率成像技术的临床需求至关重要。两光子荧光（TPF）和第二个谐波产生（SHG）显微镜是一种强大的技术，可通过在亚细胞分辨率下提供有关生物组织的结构和生化/代谢信息，而无需组织去除或外部荧光剂。但是，由于缺乏微型技术平台，其体内临床应用仍然极为限制。该多学科提案的目的是开发全纤维扫描内部显微镜技术，该技术能够将TPF/SHG显微镜带到诊所进行内部器官成像。它涉及5个来自学术界的2个合作伙伴，三个来自行业。 The proposed technology will integrate all essential functions of a scanning laser microscope into a single flexible fiber-optic probe of a small diameter (~2.4-3.4 mm), with built-in mechanisms for femtosecond pulse delivery, dispersion management, nonlinear effect suppression, beam focusing, rapid 2D raster beam scanning, TPF/SHG collection, and focus tracking (or depth scanning).小尺寸允许其与标准的红色标记技术（例如胃镜）集成。在此提案中，我们计划应对开发这种内部细胞镜技术的主要挑战，并评估其对亚细胞分辨率成像和癌症检测的可行性。具体目的是：（1）开发新的双层纤维（DCF），纯二氧化硅核心，较大的内部外壳和数值孔径，以极大地抑制纤维内的TPF/SHG背景（例如，通过50倍）（例如50倍）并提高TPF/SHG收集效率（例如，通过〜15倍）超过了〜15倍（例如，〜15倍），而不是〜15倍。 （2）开发直径为2.1mm和0.6 Na的超级光片，以提高TPF/SHG收集效率的效率至少20倍； （3）探索一种基于高阶模式DCF的新方法，以抑制光纤中的非线性效应并提高TPF/SHG激发概率； （4）开发小占地面积（1.6 x 1.6毫米）和极低的驱动电压（10V最大值）的新型MEMS扫描仪，以实现快速的2D光栅束扫描和实时焦点跟踪。将开发具有定制DCF，Microlens和MEMS扫描仪的完全集成的内分物显微镜，能够使用3D TPF/SHG成像； （5）在体内内窥镜TPF/SHG成像中进行猪食道的成像，以评估扫描探针的性能以及设计，工程和操作问题； （6）评估拟议技术对癌症检测和肿瘤边缘鉴定的可行性，并使用体内人类食管标本，并将TPF/SHG内分镜图像与相应的组织学相关联。拟议的研究的重要性是将强大的TPF/SHG显微镜技术转化为内部器官中癌症检测和图像引导活检的临床实践，并使在原位组织组织病理学的无创实时可视化以显着提高诊断和活检的产量。此外，拟议的内分物显微镜技术也将适用于许多其他临床情况，例如手术干预的指导和生物代谢功能的体内评估。 公共卫生相关性：这项多学科建议的目的是开发和测试全纤维 - 光检查3D扫描内分镜技术，该技术可以将强大的台式TPF/SHG显微镜技术带入临床实践，以对内部器官进行成像（例如胃肠道），而不是以前使用TPF/SHG Microscophy使用。提出的技术可以通过在亚细胞分辨率下提供有关生物组织的结构和生化/代谢信息，而无需去除组织或外部荧光剂，可以以“光学活检”的形式发挥作用。拟议研究的成功完成将为我们带来一种新工具，该工具可以可视化组织病理学的原位，并提高了我们早期癌症检测，肿瘤边缘鉴定以及活检和干预措施的指导的能力。