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标本的长期，高含量，三维“孵化器”成像的需求，例如在紧密环境条件下发生的疾病进展或工程替代组织的三维（3D）器官模型。考虑到这些类型的组织构建体，我们建议设计和构建紧凑且廉价的重新复制多色共聚焦成像系统，该系统在标准的CO2或CO2/O2控制的孵化器中运行，以允许在XY阶段在多个位置收集几天或几周的3D图像数据。我们将通过对旨在概括肿瘤诱导的内皮发芽的微型体外癌症模型系统进行扩展成像实验来评估和验证仪器的独特功能。与2D模型相比，三维体外培养物可以更好地概括生理微环境，从而在类似体内的系统中提供了高度可控制的实验参数。这些类型的实验系统是细胞的较简单的2D单层和更复杂的动物成像实验之间的重要中间立场，并且是从此处描述的新方法中受益最大的研究类型的一个很好的例子。当前，对潜在的细胞和分子现象的分析通常仅限于样本数量低和单个时间点测量值。使用该项目中开发的仪器，我们将表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疗效进行长期，连续研究。到资金期结束时，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被传播到整个生物医学界。除了我们在这里提出的验证研究之外，这种类型的工具还可以在许多生物医学研究领域找到重要用途。此处描述的仪器设计将成为长期高分辨率观察的关键研究工具，对疾病进展，药物疗效研究和工程替代组织（例如软骨或心脏价值）的发展，这将导致对疾病的了解并改善人类健康。