Microfluidic platforms for measuring chemical phenotypes

这个发现计划将开发新的工具，将细胞分泌的单细胞测量化学物质与它们的基因组和/或转录组联系起来。这项研究计划的第一步是开发微流控工具，用于观察细胞群体并恢复单个或亚细胞群体以进行测序分析。为了做到这一点，我们将使用微流控技术，这种技术可以精确地操纵少量液体，并与显微镜兼容。该程序中使用的微流控设备将在阵列中放置单个细胞，以便很容易观察到它们。此外，我们将探索制造这些设备的可获得的快速原型方法，以便其他研究人员可以在他们自己的研究中实现这些设备。我们将实施化学策略，在光活化交联剂的基础上选择感兴趣的细胞。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，如微流控设备或磁性颗粒上。然后我们可以回收感兴趣的细胞，并将它们送去进行测序分析。我们将利用这个平台来理解某些基因在纤毛形成中的作用。这将有助于阐明纤毛疾病的潜在机制--一种不同类别的遗传性疾病。这项研究计划的第二步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控设备，以使用电化学来监测特定分析物的分泌。首先，我们将开发一种在微流控器件中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，这种材料可以相对容易和高通量地与微流控设备结合。这些电极将被集成在小的甜甜圈形状的阀门中，这些阀门可以捕获和隔离单个细胞。然后，我们将使用这些微流控设备来研究经过基因改造的酵母，以生产具有重要药用价值的分子。我们将能够识别有效地合成感兴趣的分子的细胞，我们可以回收细胞进行测序分析，并确定进行了哪些基因修改。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来说很重要。此外，这里开发的工具可以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。

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.