ADVANCED MICROFLUIDIC DEVICES FOR CELL/MOLECULAR BIOLOGY

描述(改编自申请人的摘要)：先进微流控技术 将发展能力，以解决广泛的生化反应 将受益于精确和自动化的纳米到的测量问题 具有高串行吞吐能力的亚纳升级别的操作。 这些设备还将使自己成为大规模并行扩展的领先者 为生物化学的产生提供更大的生产能力 信息。这项工作的重点将是开发 可操纵亚纳升生化的微细加工沟道器件 以可控的方式反应体积以产生实验结果 每个通道的速率为1-10赫兹。这些反应体积将能够 含有分子或颗粒物种，没有扩散损失。这个 这些反应体积中的颗粒物种类的占有量，例如， 生物细胞或组合库珠，将从 单个实体到多个实体。因此，高度自动化的单电池 将有可能使用这些设备以及合奏进行调查 研究，即一组细胞的反应。个人 反应体积将以大致相同的方式进行连续操作 作为数字移位寄存器，允许对每个反应进行唯一标识 “细胞”贯穿整个实验。这些反应体积移位寄存器设备 将与向单个反应中添加试剂的功能相集成 在“试剂站”的体积，然后提取材料进行分析 在“化验站”用光学手段直接测定反应体积将 也是有可能的。拟议能力的发展将使 能够进行多种不同类型的高通量实验 (105-106/天)使用由PC计算机尺寸驱动的单个微流控设备 硬件。可以通过以下方式按比例实现更大的吞吐量 以最低限度增加硬件需求的并行化。这项技术 将应用于筛选分子或细胞等问题 使用分离/组合文库中的单个珠子的目标，筛选 用于RNA或蛋白质表达的单个细胞，遗传诊断筛查 单细胞水平或执行单细胞信号转导研究。

DESCRIPTION (Adapted from the applicant's abstract): Advanced microfluidic capabilities will be developed to address a broad range of biochemical measurement problems that will benefit from precise and automated nano- to subnanoliter scale manipulations with high serial throughput capability. These devices will also lend themselves to massive parallel expansion leading to greater throughput possibilities for the generation of biochemical information. The focus of this work will be the development of microfabricated channel devices that can manipulate subnanoliter biochemical reaction volumes in a controlled manner so as to produce experimental results at rates of 1-10 Hz per channel. These reaction volumes will be capable of containing molecular or particulate species without diffusive losses. The occupation number of particulate species in these reaction volumes, e.g., biological cells or combinatorial library beads, will be controllable from single to multiple entities. Thus, highly automated single cell investigations will be possible with these devices as well as ensemble studies, i.e., the response from a collection of cells. The individual reaction volumes will be manipulated in serial fashion, in much the same way as a digital shift register, allowing unique identification of each reaction "cell" throughout an experiment. These reaction volume shift register devices will be integrated with capabilities to add reagents to individual reaction volumes at "reagent stations" and subsequently extract material for analysis at "assay stations." Direct assay of reaction volumes by optical means will also be possible. Development of the proposed capabilities will enable the ability to perform a number of different types of high throughput experiments (105-106/day) using single microfluidic devices driven by PC computer size hardware. Proportionately greater throughput can be achieved by parallelization with minimally increased hardware demands. This technology will have application to problems such as screening molecular or cellular targets using single beads from split/pool combinatorial libraries, screening single cells for RNA or protein expression, genetic diagnostic screening at the single cell level or performing single cell signal transduction studies.