Targeted Multi-Spectral Dual Axes Confocal Imaging of In Vivo Molecular Expressio

体内分子表达的靶向多光谱双轴共焦成像

• 批准号: 8410480
• 负责人: Katsuo Kurabayashi
• 金额: $ 45.01万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-25 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8410480
• 关键词: Affinity; Animal Model; Animals; Architecture; Behavior; Binding; Biological Phenomena; Biomedical Engineering; Breast; Breast Cancer Cell; Cancer Biology; Cell Communication; Cell surface; Cells; Chest; Collection; Colon; Colon Carcinoma; Colon Hyperplastic Polyp; Colorectal Cancer; Detection; Development; Devices; Dimensions; Distal; Diversity Library; Duct (organ) structure; Dyes; Dysplasia; Electronics; Engineering; Epithelial; Epithelium; Film; Fluorescence; Genetically Engineered Mouse; Goals; Hyperplasia; Image; Implant; Investigation; Label; Lateral; Lead; Life; Ligands; Light; Lighting; Longitudinal Studies; Malignant Neoplasms; Michigan; Microscope; Miniaturization; Molecular; Morphologic artifacts; Motion; Mucous Membrane; Mus; Neoplasms; Normal tissue morphology; Optics; Pattern; Penetration; Peptides; Performance; Phage Display; Process; Public Health; Research; Research Personnel; Resolution; Scanning; Source; Speed; Surface; Techniques; Thick; Time; Tissues; Translating; Tumor Angiogenesis; Universities; Vascular Endothelium; Work; Xenograft Model; Xenograft procedure; cancer imaging; chemokine receptor; college; design; detector; drug discovery; flexibility; fluorescence imaging; fluorescence microscope; in vivo; innovation; instrument; malignant breast neoplasm; medical schools; mouse model; novel; optical imaging; prototype; public health relevance; receptor

DESCRIPTION (provided by applicant): Targeted Multi-Spectral Dual Axes Confocal Imaging of In Vivo Molecular Expression The broad, long-term objective of this BRP is to develop a miniature intra-vital imaging instrument to study molecular mechanisms of cancer biology in the epithelium of live animals. This novel optical design uses off-axis illumination and collection of light to produce superior dynamic range for collecting vertical cross-sectional fluorescence images. We will assemble an inter-disciplinary team of investigators from the College of Engineering and the School of Medicine at the University of Michigan. The specific aims are 1) to develop a scanning mechanism to achieve real time, multi- spectral vertical cross-sectional imaging, 2) to select affinity peptides that bind to dysplasia in colonic mucosal epithelium and to CXCR7 in breast vascular endothelium, and 3) to validate imaging performance in live animal models. The dual axes confocal architecture uses the overlapping focus of two separate low numerical aperture objectives to achieve sub-cellular resolution and long working distance. Post-objective scanning provides a large field-of-view as well as scalability and miniaturization of the optics, and a replicated parabolic mirror directs collimated, multi-spectral beams to a common focus. This unique design can view the epithelial differentiation pattern in the basilar to luminal direction. Real time imaging of the epithelium will be achieved by developing a vertical actuator that uses thin film piezoelectric materials that have the size, speed, force, and linearity to overcome motion artifact (>10 frames/sec). This device will be combined with a lateral scanning micro-mirror that is driven by parametric resonance. Affinity peptides will be selected using the technique of phage display by biopanning a high diversity library against cells and tissues that over express surface targets associated with neoplasia. Multiple probes can be near-infrared labeled to achieve the tissue penetration depths expected (>500 microns). Imaging performance will be validated by inserting the miniature prototype into the colon to evaluate the epithelium of the distal mucosa or by placing against the chest to image the epithelium of breast ducts of live animals. Public Health: This general purpose instrument can be used to study molecular processes in the epithelium of live animal models with sub-cellular resolution, high dynamic range, and full imaging depth, allowing for longitudinal investigation of mechanisms of cancer biology and providing a new platform for drug discovery.

描述（由申请人提供）：活体分子表达的靶向多光谱双轴共聚焦成像本BRP的广泛和长期目标是开发一种微型的活体内成像仪器，以研究活体动物上皮中癌症生物学的分子机制。这种新颖的光学设计使用离轴照明和光收集来产生优越的动态范围，用于收集垂直横截面荧光图像。我们将组建一个由密歇根大学工程学院和医学院组成的跨学科研究小组。具体目的是：1)开发一种扫描机制，实现实时、多光谱垂直横断面成像；2)选择与结肠黏膜上皮发育不良和乳腺血管内皮CXCR7结合的亲和肽；3)验证活体动物模型的成像性能。双轴共焦结构利用两个独立的低数值孔径物镜的重叠聚焦来实现亚细胞分辨率和较长的工作距离。后物镜扫描提供了一个大的视场，以及光学器件的可扩展性和小型化，一个复制的抛物面镜将准直的多光谱光束引导到一个共同的焦点。这种独特的设计可以观察基底向管腔方向的上皮分化模式。上皮的实时成像将通过开发一种垂直致动器来实现，该致动器使用薄膜压电材料，具有克服运动伪影（bbb10帧/秒）的尺寸、速度、力和线性。该装置将与一个由参数共振驱动的横向扫描微镜相结合。利用噬菌体展示技术，对与肿瘤相关的细胞和组织进行生物筛选，筛选高多样性文库中的亲和肽。多个探针可以近红外标记，以达到预期的组织穿透深度（bbb500微米）。成像性能将通过将微型原型插入结肠以评估远端粘膜上皮或将其放置在胸部对活体动物的乳腺导管上皮进行成像来验证。公共卫生：该通用仪器可用于研究活体动物模型上皮的分子过程，具有亚细胞分辨率、高动态范围和全成像深度，允许对癌症生物学机制进行纵向研究，并为药物发现提供新的平台。