Developing an automated yeast dissection system for aging research

开发用于衰老研究的自动化酵母解剖系统

• 批准号: 9463844
• 负责人: Weiwei Dang
• 金额: $ 22.5万
• 依托单位: INNOVATIVE BIOCHIPS, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2018-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9463844
• 关键词: Agar; Age; Age of Onset; Aging; Aging-Related Process; Automation; Award; Biological Assay; Biological Sciences; Cell Aging; Cells; Chimeric Proteins; Computer software; Data; Data Collection; Disease; Dissection; Environment; Enzymes; Eukaryota; Eukaryotic Cell; Gene Expression; Genes; Goals; Image; Image Analysis; Imagery; Impairment; Incubators; Individual; Legal patent; Longevity; Measurement; Medicine; Methods; Microdissection; Microfluidics; Modeling; Molecular; Morphology; Mothers; Organelles; Pathway interactions; Phase; Phenotype; Polymers; Procedures; Proteins; Public Health; Publishing; Research; Research Personnel; Resolution; Risk Factors; Saccharomyces cerevisiae; Saccharomycetales; Signal Transduction; Silicon; Study models; System; Technology; Thick; Time; Yeasts; aged; base; biochip; college; cost; daughter cell; design; experimental study; fluorescence imaging; innovation; mortality; mutant; novel; organic base; programs; prototype; spatiotemporal; yeast genetics

Project Summary Developing an Automated Yeast Dissection System for Aging Research Aging is the single greatest risk factor for diseases that are principal causes of mortality. The objectives of aging research are to discover key genes and pathways related to aging that may eventually contribute to retardation of aging and a delay in the onset of age-associated diseases. The budding yeast Saccharomyces cerevisiae has been a powerful model for the study of aging and has enabled significant contributions to our understanding of basic mechanisms of aging in eukaryotic cells. However, traditional assays of yeast aging, including microdissection methods, have technical challenges; for instance, the methods are low-throughput and the experimental procedures are laborious. An experiment typically lasts several weeks or months, and requires overnight storage of the assayed cells at a refrigerator to pause replication throughout the course of experiment. This tedious procedure has substantially hindered progress in the field of aging research. We have developed an innovative and highly effective microfluidic platform for continuous and automatic dissection of daughter cells without disturbing mother cells as they bud. The platform allows an automated whole-lifespan tracking with high spatiotemporal resolution and large-scale quantification of single yeast cells, resulting in significant reduction of labor, time, and cost. In addition, the high-resolution florescence imaging of yeast cells grown in constant and dynamically changing environments offers the ability to examine the dynamics of gene expression and signaling networks in a high-throughput manner. The quantity and types of data acquired by this platform are impossible with the traditional assay methods. Herein, we propose to develop automated dissection platform that enable fast and high-throughput studies of yeast aging. We will validate the reliability and capability of the proposed platform We propose to validate the prototype automated dissection platform for two important assays in yeast aging research: determination of replicative lifespan for mutants in an automated and unattended fashion, and continuous tracking the changes in abundance and cellular localization for specific GFP fusion proteins during the entire aging process. Longer term plan of this proposal is to commercialize a fully automation benchtop system for high-throughput studies of cellular aging.

项目摘要 开发一种自动化的酵母解剖系统，以进行衰老研究 衰老是疾病的最大危险因素，这是死亡主要原因。目标的目标 衰老的研究是发现与衰老有关的关键基因和途径，这可能最终有助于 衰老的迟缓和与年龄相关疾病发作的延迟。萌芽的酵母菌 酿酒酵母一直是研究衰老的强大模型，并为我们的 了解真核细胞中衰老的基本机制。但是，传统的酵母老化测定法， 包括微分解方法，面临技术挑战；例如，这些方法是低通量的， 实验程序很费力。一个实验通常持续数周或几个月，需要 在整个实验过程中，在冰箱中隔夜存储测定的细胞以暂停复制。 这种乏味的程序极大地阻碍了老化研究领域的进步。我们已经发展了 一个创新且高效的微流体平台，用于连续和自动解剖子细胞 芽时不会干扰母细胞。该平台允许自动化全杆台跟踪，高 单个酵母细胞的时空分辨率和大规模定量，导致显着降低 劳动，时间和成本。另外，在恒定和 动态变化的环境提供了检查基因表达和信号传导动力学的能力 网络以高通量方式。该平台获取的数据数量和类型是不可能的 使用传统的测定方法。在此，我们建议开发自动解剖平台 酵母老化的快速和高通量研究。我们将验证提议的可靠性和能力 平台我们建议验证原型自动解剖平台，以在酵母中进行两个重要测定 衰老研究：以自动和无人值守的方式确定突变体的复制寿命，以及 连续跟踪特定GFP融合蛋白的丰度和细胞定位的变化 整个老化过程。该提案的长期计划是商业化完全自动化的台式机 用于细胞衰老的高通量研究的系统。