Microfluidic platforms for measuring chemical phenotypes

用于测量化学表型的微流体平台

• 批准号: RGPIN-2022-03797
• 负责人: Rackus, Darius
• 金额: $ 1.82万
• 依托单位: Ryerson University
• 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=748544
• 关键词: Microfluidic; platforms; measuring; chemical; phenotypes

This Discovery program will develop new tools to link single cell measurements chemicals secreted by cells with their genome and/or transcriptome. The first stream of this research program seeks to develop microfluidic tools for observing populations of cells and recovering single or sub-populations of cells for sequencing analysis. To do this, we will use microfluidic technologies, which can manipulate small volumes of liquid with precision and are compatible with microscopy. The microfluidic devices used in this program will place single cells within an array so they can be easily observed. Moreover, we will explore accessible rapid-prototyping methods for fabricating these devices so that other researchers can implement these devices in their own research. We will implement chemical strategies for selecting cells of interest based off of photo-activated crosslinking. By shining light onto a targeted cell, we can chemically link it onto a surface, such as the microfluidic device or onto a magnetic particle. We can then recover cells of interest and send them for sequencing analysis. We will apply this platform to understanding the role of certain genes in cilia formation. This will help elucidate underlying mechanisms of ciliopathies-a diverse category of genetic disorders. The second stream of this research program will develop tools to measure chemicals secreted by single cells. We aim to develop microfluidic devices with arrays of hundreds of electrodes to monitor secretion of specific analytes using electrochemistry. First, we will develop a fabrication method for integrating high-density electrode arrays within microfluidic devices. Our method will use conductive materials that can readily be incorporated with the microfluidic device with relative ease and high-throughput. These electrodes will be integrated within small donut-shaped valves that can trap and isolate single cells. We will then use these microfluidic devices to study yeast that have been genetically modified to produce a molecule of pharmaceutical importance. We will be able to identify cells that efficiently synthesize a molecule of interest and we can recover the cells for sequencing analysis and determine which genetic modifications were made. The results of this research program have impact in areas such as disease and biomanufacturing, two areas of importance to Canadians. Further, the tools developed here can be applied to answer both basic and applied research questions in the areas of cell biology, functional genomics, and personalized healthcare.

该发现计划将开发新的工具，将单细胞测量细胞分泌的化学物质与其基因组和/或转录组联系起来。该研究计划的第一个流旨在开发微流体工具，用于观察细胞群并回收单个或亚群细胞进行测序分析。为此，我们将使用微流体技术，该技术可以精确地操纵小体积的液体，并与显微镜兼容。该计划中使用的微流体装置将单个细胞放置在阵列中，因此可以很容易地观察它们。此外，我们将探索制造这些设备的快速原型方法，以便其他研究人员可以在自己的研究中实现这些设备。我们将实施基于光活化交联的化学策略来选择感兴趣的细胞。通过将光照射到目标细胞上，我们可以将其化学连接到表面上，例如微流体装置或磁性颗粒上。然后，我们可以回收感兴趣的细胞并将其送去测序分析。我们将应用这个平台来了解某些基因在纤毛形成中的作用。这将有助于阐明潜在的机制，睫状体病-一个不同类别的遗传性疾病。该研究计划的第二部分将开发测量单细胞分泌的化学物质的工具。我们的目标是开发具有数百个电极阵列的微流体装置，以使用电化学来监测特定分析物的分泌。首先，我们将开发一种用于在微流体装置内集成高密度电极阵列的制造方法。我们的方法将使用导电材料，这些材料可以相对容易和高通量地与微流体装置结合。这些电极将被集成在小型甜甜圈形阀门中，可以捕获和隔离单细胞。然后，我们将使用这些微流控装置来研究经过遗传修饰的酵母，以产生具有药物重要性的分子。我们将能够识别有效合成感兴趣分子的细胞，我们可以回收细胞进行测序分析，并确定进行了哪些遗传修饰。这项研究计划的结果在疾病和生物制造等领域产生了影响，这两个领域对加拿大人很重要。此外，这里开发的工具可以应用于回答细胞生物学，功能基因组学和个性化医疗保健领域的基础和应用研究问题。