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

描述（由申请人提供）：本项目解决了对多层标本长期、高含量、三维“培养箱内”成像的新兴需求，如疾病进展的三维器官模型或在严格环境条件下发育的工程替代组织。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廉价的多色共聚焦成像系统，该系统在标准CO2或CO2/O2控制的培养箱内运行，允许在XY台上的多个位置收集数天或数周的3D图像数据。我们将评估和验证我们的仪器的独特功能，通过在一个微制造的体外癌症模型系统上进行扩展的成像实验，该系统旨在重现肿瘤诱导的内皮发芽。三维体外培养已被证明比二维模型更好地再现生理微环境，在更类似于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地带，并且是将从这里描述的新方法中获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局限于低样本数量和单时间点测量。使用本项目开发的仪器，我们将展示我们可以进行肿瘤血管生成、细胞运动和相互作用以及抗血管生成治疗效果的长期、连续研究。在融资期结束时，我们将创建并演示一个新的成像平台，该平台将为商业开发做好准备，这样我们设计的独特技术就可以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，这种类型的仪器将在生物医学研究的许多领域中找到重要的用途。这里描述的仪器设计将成为疾病进展模型系统的长期高分辨率观察，药物功效研究和工程替代组织（如软骨或心脏价值）开发的关键研究工具，这将导致对疾病的更好理解并有助于改善人类健康。

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.