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ADVANCED MICROFLUIDIC DEVICES FOR CELL/MOLECULAR BIOLOGY

用于细胞/分子生物学的先进微流体装置

基本信息

批准号：
6188621
负责人：
JOHN Michael RAMSEY
金额：
$ 51.15万
依托单位：
LOCKHEED MARTIN ENERGY RESEARCH CORP
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-09-30 至 2002-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6188621
关键词：
biomedical equipment development cell biology electrophoresis fluid flow micromanipulator molecular biology single cell analysis

项目摘要

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.

描述（改编自申请人的摘要）：先进的微流体将开发能力来解决广泛的生化问题测量问题将受益于精确和自动化的纳米技术具有高串行吞吐量能力的亚纳升规模操作。这些设备还将有助于大规模并行扩展产生生化物质的更大通量可能性信息。这项工作的重点将是开发可以操纵亚纳升生物化学的微加工通道装置以受控方式调节反应体积以产生实验结果每个通道的速率为 1-10 Hz。这些反应体积将能够含有分子或颗粒物质，没有扩散损失。这这些反应体积中颗粒物的占据数量，例如，生物细胞或组合库珠，将由单个到多个实体。因此，高度自动化的单细胞使用这些设备和整体可以进行调查研究，即一组细胞的反应。个人反应体积将以串行方式操作，方式大致相同作为数字移位寄存器，允许每个反应的唯一标识整个实验过程中的“细胞”。这些反应体积移位寄存器装置将与向单个反应添加试剂的功能集成 “试剂站”的体积，随后提取材料进行分析在“化验站”。通过光学手段直接测定反应体积将也有可能。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体装置硬件。可以通过以下方式实现成比例更大的吞吐量并行化，同时最大限度地减少硬件需求的增加。这项技术将应用于筛选分子或细胞等问题使用来自分割/池组合文库的单个珠子进行靶向，筛选用于 RNA 或蛋白质表达的单细胞，遗传诊断筛选单细胞水平或进行单细胞信号转导研究。