Reducing morbidity in surgical resections: Third-harmonic generation microscopy a

降低手术切除的发病率：三次谐波发生显微镜a

• 批准号: 8720770
• 负责人: CHRIS B SCHAFFER
• 金额: $ 23.43万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8720770
• 关键词: Accidental Injury; Animal Model; Biological Assay; Cellular Structures; Cytoskeleton; Data; Development; Device or Instrument Development; Dyes; Effectiveness; Endoscopes; Environment; Erectile dysfunction; Excision; Fiber; Fluorescence; Generations; Goals; Grant; Human; Image; Imagery; Imaging Device; Imaging Techniques; Implant; Injury; Label; Laparoscopic Surgical Procedures; Lasers; Lead; Light; Malignant neoplasm of prostate; Maps; Methods; Microscope; Microscopy; Modality; Modeling; Morbidity - disease rate; Mus; Myelin; Nerve; Nerve Fibers; Neuraxis; Operating Rooms; Operative Surgical Procedures; Optics; Outcome; Patients; Peripheral Nerves; Prostate; Prostatectomy; Proteins; Publishing; Research; Rodent; Sensitivity and Specificity; Signal Transduction; Site; Solutions; Spinal Cord; Surgeon; Surgical complication; System; Testing; Time; Tissues; Visual Fields; Work; base; design; efficacy testing; fluorescence imaging; imaging modality; improved; in vivo imaging; instrument; neoplastic cell; nerve injury; novel; novel strategies; operation; optical imaging; pre-clinical; prevent; prostate cancer model; prototype; public health relevance; research study; second harmonic; success; tool; tumor; two-photon

DESCRIPTION (provided by applicant): Unintentional injury to nerves is one of the primary causes of morbidity during surgical resection of tumors as well as in other surgeries. Nerve fibers are difficult to distinguish from the surrounding tissues during surgery, making it difficul for surgeons to carefully work around and avoid their injury. Broadly speaking, the goal of the proposed work is to establish third-harmonic generation (THG) microscopy as an approach to visualize, with high sensitivity, nerve fibers intraoperatively. We have previously shown that THG provides a high-sensitivity approach to imaging myelin in the central nervous system of rodents and have used this for in vivo imaging of myelin in the spinal cord. We recently collected preliminary data that shows this approach can highlight peripheral nerves as well. Ultimately, this imaging modality could allow the surgeon to visualize nerve fibers and then work carefully near them, thereby preventing surgical complications associated with nerve injury. We identify three Specific Aims necessary to complete this work. First, we build on our preliminary data and identify the best set of laser and imaging parameters to visualize myelinated peripheral nerve bundles with THG imaging. Second, we assess the effectiveness of THG imaging to spare nerves by comparing the outcome of prostate resection surgeries in rodents done with and without this novel imaging modality. In this Aim, we further build on previous results and examine not just how well nerves are spared when visualized with THG, but also how completely tumors are extracted when they are imaged with two-photon excited fluorescence (2PEF) microscopy. Critically, the laser and imaging systems necessary for THG imaging are completely compatible with 2PEF imaging, allowing these two modalities to work in concert to help a surgeon both maximize tumor extraction and minimize nerve damage. Taken together, these two aims will establish the utility of our approach to nerve imaging, but use in an operating room environment will clearly require development of a less-unwieldy instrument than our current microscopes. Toward this end, in the third Aim, we design an endoscope that is optimized for THG (as well as 2PEF) imaging and could be used in laparoscopic surgeries (such as many prostate cancer resections) in humans. Taken together, this work will establish the utility of THG as a viable imaging modality for use in surgery and will develop the first-generation instrument that could be used in an operating room. If successful, this work could dramatically reduce the post-surgical morbidity of resection surgeries - especially in cases where critical nerves are close to the resection site, such as in the prostate - and therefore improve patient outcomes.

描述（由申请人提供）：无意的神经损伤是肿瘤手术切除以及其他手术期间发病的主要原因之一。在手术过程中，神经纤维很难与周围组织区分开来，这使得外科医生很难仔细工作并避免受伤。从广义上讲，拟议工作的目标是建立三次谐波发生（THG）显微镜作为一种在术中以高灵敏度可视化神经纤维的方法。我们之前已经证明，THG 提供了一种高灵敏度的方法来对啮齿动物中枢神经系统中的髓磷脂进行成像，并将其用于脊髓中髓磷脂的体内成像。我们最近收集的初步数据表明这种方法也可以突出周围神经。最终，这种成像方式可以让外科医生可视化神经纤维，然后在它们附近仔细工作，从而防止与神经损伤相关的手术并发症。我们确定了完成这项工作所需的三个具体目标。首先，我们以初步数据为基础，确定最佳的激光和成像参数集，以通过 THG 成像可视化有髓鞘周围神经束。其次，我们通过比较使用和不使用这种新型成像方式的啮齿动物前列腺切除手术的结果，评估了 THG 成像对保护神经的有效性。在这个目标中，我们进一步建立在以前的结果的基础上，不仅检查用 THG 可视化时神经的完好程度，而且还检查用双光子激发荧光 (2PEF) 显微镜成像时肿瘤被提取的完全程度。至关重要的是，THG 成像所需的激光和成像系统与 2PEF 成像完全兼容，使这两种模式协同工作，帮助外科医生最大限度地提取肿瘤并最大限度地减少神经损伤。总而言之，这两个目标将确立我们的神经成像方法的实用性，但在手术中使用 房间环境显然需要开发一种比我们当前的显微镜更方便的仪器。为此，在第三个目标中，我们设计了一种针对 THG（以及 2PEF）成像进行优化的内窥镜，可用于人类腹腔镜手术（例如许多前列腺癌切除术）。总而言之，这项工作将确立 THG 作为手术中可行的成像方式的实用性，并将开发可在手术室使用的第一代仪器。如果成功，这项工作可以显着降低切除手术的术后发病率，特别是在关键神经靠近切除部位的情况下，例如前列腺，从而改善患者的治疗效果。