Methods development for the measurement of single-cell molecular state

单细胞分子状态测量方法的开发

• 批准号: RGPIN-2020-05254
• 负责人: Knapp, David
• 金额: $ 2.7万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=757499
• 关键词: Methods; development; measurement; single; cell

Tissue systems are made up of a complex mixture of cells each with unique behaviours and regulatory mechanisms controlling them. Interestingly, even when purified groups of these cells with theoretically the same roles have been examined closely, they too have been shown to have a mix of individual behaviours/regulation. Such complexity is likely important in maintaining proper balance in the tissue system and for adaptation to changing conditions. Despite this, most common techniques used to measure how cells function are only able to measure the population average. While these bulk techniques can give extremely useful benchmarks of the general characteristics of a group of cells, they necessarily ignore much of the regulatory complexity which control the system behaviour. Techniques exist to look at some of the control mechanisms of single cells, however, they tend to be restricted to one type of control where cells use multiple mechanisms to ensure proper regulation. These techniques also tend to be both difficult to perform and expensive. We will develop new ways to measure multiple mechanisms of cell regulation at the same time in single cells to give a more complete picture of their decision-making process. To facilitate the use of these techniques we will also develop automated ways of performing these protocols while decreasing costs by reducing the scale of reactions down to sub-microliter volumes. Finally, we will apply nano-scale electronic detectors to create a new method for measuring proteins in single cells. Overall, the proposed program will provide valuable fundamental tools which will be applicable to a wide range of basic cell biological research.

组织系统由复杂的细胞混合物组成，每个细胞都有独特的行为和控制它们的调节机制。有趣的是，即使在理论上具有相同作用的这些细胞的纯化组被仔细检查时，它们也被证明具有个体行为/调节的混合。这种复杂性在维持组织系统的适当平衡和适应变化的条件方面可能是重要的。尽管如此，大多数用于测量细胞功能的常用技术只能测量群体平均值。虽然这些批量技术可以提供一组细胞的一般特征的非常有用的基准，但它们必然忽略了控制系统行为的许多监管复杂性。现有技术可以观察单细胞的一些控制机制，然而，它们往往限于一种类型的控制，其中细胞使用多种机制来确保适当的调节。这些技术也往往既难以执行又昂贵。我们将开发新的方法来同时测量单个细胞中的多种细胞调节机制，以更全面地了解它们的决策过程。为了促进这些技术的使用，我们还将开发执行这些协议的自动化方法，同时通过将反应规模降低到亚微升体积来降低成本。最后，我们将应用纳米级电子探测器来创建一种测量单细胞中蛋白质的新方法。总的来说，拟议的计划将提供有价值的基本工具，将适用于广泛的基础细胞生物学研究。