Multiplexed Ovarian Cancer Microfluidic Tissue Microarray

项目摘要 我们提出了一个自动化的、基于细胞和组织的多路传输的仪器系统 实验(即组织微阵列)称为微流控流动细胞阵列(MFCA)。MFCA 由集成有倒置荧光显微镜的微流控流动细胞阵列组成， 允许同时观察48个流动室或更高的单个流动室 放大。建议的仪器将用于新的基于细胞和组织的分析在 高度并行的方式，否则很难执行。我们的目标是为 生产一种组织微阵列，它保持了体内细胞的许多特征 并允许研究细胞表面、细胞-细胞，特别是肿瘤间质 以多重方式进行相互作用、细胞毒性研究和组织毒性研究。作为一个阶段 我的申请，这项建议的重点是卵巢癌组织切片毒性测试，以确定 针对特定患者的最佳化疗药物(即基于 组织反应)；然而，可以设想卵巢癌以外的多种应用 包括：组合细胞刺激、毒性或治疗分析、干细胞筛选和 分化控制研究，多重原代细胞分析，生物材料筛选，剪切法 应激和细胞黏附研究，以及组合免疫组织化学。这个平台 方法和与显微镜的简单集成将允许采用和适应 世界各地的研究人员。 确定了以下假设和具体目标，以证明 微流控流动细胞阵列技术与倒置荧光显微镜的集成 基于细胞和组织的分析。 具体目标1.翻译现有组织培养方法的终点分析，包括 用四甲基偶氮唑盐比色法和数字病理学对MFCA进行检测。 具体目标2.设计MFCA打印头，将打印头对接的影响降至最低 组织培养切片。 具体目标3.优化不同治疗剂在MFCA通道中的药物输送 应用于小鼠卵巢组织切片，并与现有的组织培养方法进行比较。

Project Summary We propose an instrument system for automated, multiplexed cell- and tissue-based experiments (i.e. tissue microarrays) called the Microfluidic Flow Cell Array (MFCA). The MFCA consists of a microfluidic flow cell array integrated with an inverted fluorescent microscope, allowing the observation of 48 flow chambers simultaneously or an individual chamber at higher magnification. The proposed instrument will be used to novel cell- and tissue-based assays in a highly parallel manner that are otherwise difficult to perform. Our goal is to build a platform for producing a tissue microarray that maintains many of the characteristics found in cells in vivo and allows for the investigation of cell-surface, cell-cell, and especially tumor-stroma interactions, cell toxicity studies, and tissue toxicity studies in a multiplexed fashion. As a Phase I application, this proposal's focus is ovarian cancer tissue slice toxicity testing to determine the optimal chemotherapeutic agents for a specific patient (i.e. personalized treatment based on the tissue reaction); however, multiple applications can be envisioned beyond ovarian cancer including: combinatorial cell stimulation, toxicity, or therapeutic assays, stem cell screening and differentiation control studies, multiplexed primary cell assays, biomaterials screening, shear stress and cell adhesion studies, and combinatorial immunohistochemistry. This platform approach and simple integration with a microscope will allow for adoption and adaptability for researchers worldwide. The following hypotheses and specific aims have been identified to prove the feasibility of integrating the microfluidic flow cell array technology with an inverted fluorescent microscope for cell and tissue based assays. Specific Aim 1. Translate end point assays for existing tissue culture approaches including the MTT Assay and digital pathology to the MFCA. Specific Aim 2. Engineer the MFCA printhead to minimize impact of the printhead docking onto the tissue culture slice. Specific Aim 3. Optimize drug delivery of different therapeutic agents in the MFCA channels using to the mouse ovarian tissue slice and compare to existing tissue culture approaches.