Microfluidic platforms for measuring chemical phenotypes

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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.