A miniature confocal for long term 3D multicolor imaging within a C02 incubator

用于 CO2 培养箱内长期 3D 多色成像的微型共焦

• 批准号: 8575684
• 负责人: WARREN R ZIPFEL
• 金额: $ 16.82万
• 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: 2013-08-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8575684
• 关键词: Address; Animals; Antineoplastic Agents; Area; Beds; Biological; Biological Models; Biomedical Research; Cancer Model; Carbon Dioxide; Cartilage; Cell Differentiation process; Cells; Color; Communities; Complex; Computer software; Confocal Microscopy; Controlled Environment; Data; Detection; Development; Devices; Disease; Disease Progression; Engineering; Environment; Fiber; Fluorescence; Funding; Goals; Growth; Health; Heart; Histocompatibility Testing; Human; Humidity; Image; In Situ; In Vitro; Incubators; Investigation; Lasers; Lead; Measurement; Mechanical Stimulation; Mechanics; Microscope; Microscopic; Microscopy; Mind; Modeling; Modification; Molecular; Molecular Analysis; Monitor; Neoplasm Metastasis; Operating System; Organ Model; Physiological; Positioning Attribute; Process; Property; Regulation; Research; Research Design; Research Infrastructure; Resolution; Role; Sampling; Scanning; Specimen; Staging; System; Technology; Testing; Three-Dimensional Image; Three-Dimensional Imaging; Time; Tissue Engineering; Tissues; Tumor Angiogenesis; Validation; angiogenesis; base; cancer therapy; cell motility; design; drug efficacy; imaging modality; improved; in vivo; instrument; monolayer; novel strategies; prototype; public health relevance; research study; scaffold; spatiotemporal; three-dimensional modeling; tool; transmission process; tumor; tumor progression; validation studies

DESCRIPTION (provided by applicant): This project addresses an emerging demand for long-term, high content, three-dimensional "intra- incubator" imaging of multilayer specimens such as three dimensional (3D) organ models of disease progression or engineered replacement tissues developing under tight environmental conditions. With these types of tissue constructs in mind, we propose to design and build a compact and inexpensive- to-replicate multi-color confocal imaging system that operates within a standard CO2 or CO2/O2 controlled incubator to allow for days or weeks of 3D image data to be collected at multiple positions on an XY stage. We will evaluate and verify our instrument's unique functionality by carrying out extended imaging experiments on a microfabricated in vitro cancer model system designed to recapitulate tumor-induced endothelial sprouting. Three-dimensional in vitro cultures have been shown to better recapitulate the physiological microenvironment than 2D models, providing highly controllable experimental parameters in a more in vivo-like system. These types of experimental systems are an important middle ground between simpler 2D monolayers of cells and more complex animal imaging experiments, and are an excellent example of the types of studies that would benefit the most from the novel approach described here. Currently, analysis of the underlying cellular and molecular phenomena is usually limited to low sample numbers and single time point measurements. Using the instrument developed in this project we will show that we can perform long-term, continuous studies of tumor angiogenesis, cellular motility and interactions, and the efficacy of anti-angiogenic therapies. By the end of the funding period we will have created and demonstrated a new imaging platform that will be ready for commercial development so that the unique technology we design is disseminated to the entire biomedical community. This type of instrument would find important uses in many areas of biomedical research in addition to the validation study we propose here. The instrument design described here will become a critical research tool for long term high resolution observations of model systems of disease progression, drug efficacy studies and development of engineered replacement tissues such as cartilage or heart values, which will lead to an improved understanding of disease and help improve human health.

描述（由申请人提供）：该项目解决了对多层样本的长期、高内涵、三维“培养箱内”成像的新兴需求，例如疾病进展的三维（3D）器官模型或开发的工程替代组织在严格的环境条件下。考虑到这些类型的组织结构，我们建议设计和构建一个紧凑且复制成本低廉的多色共焦成像系统，该系统在标准 CO2 或 CO2/O2 控制的培养箱中运行，以允许数天或数周的 3D 图像在 XY 平台上的多个位置收集数据。我们将通过在微制造的体外癌症模型系统上进行扩展成像实验来评估和验证我们仪器的独特功能，该模型旨在重现肿瘤诱导的内皮芽生长。三维体外培养物已被证明比二维模型能更好地概括生理微环境，在更像体内的系统中提供高度可控的实验参数。这些类型的实验系统是更简单的二维单层细胞和更复杂的动物成像实验之间的重要中间立场，并且是将从此处描述的新颖方法中受益最多的研究类型的一个很好的例子。目前，对潜在细胞和分子现象的分析通常仅限于低样本数量和单时间点测量。使用该项目开发的仪器，我们将证明我们可以对肿瘤血管生成、细胞运动和相互作用以及抗血管生成疗法的功效进行长期、连续的研究。在资助期结束时，我们将创建并展示一个新的成像平台，该平台将为商业开发做好准备，以便我们设计的独特技术能够传播到整个生物医学界。除了我们在此提出的验证研究之外，这种类型的仪器还将在生物医学研究的许多领域找到重要用途。这里描述的仪器设计将成为一个关键的研究工具，用于长期高分辨率观察疾病进展模型系统、药物功效研究和工程替代组织（例如软骨或心脏价值）的开发，这将导致更好地了解疾病和帮助改善人类健康。