High adhesive substrate and microfluidic system for the comet assay

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This project aims to produce an improved system for running a comet assay on chip with a far higher degree of control, uniformity and at lower costs of manufacturing than current solutions. A comet assay is a specific genomic test that looks for genetic damage done to cells after exposure to mutagenic agents. Cells are encased in hydrogels to keep them in position, exposed to conditions of interest, then ruptured and the DNA stained for fluorescence visualization. An electric field is applied and damaged DNA spreads more, providing a "comet" like appearance, while non-damaged DNA remains tightly clustered. While the comet assay is extremely useful, it is a slow, very manual process that relies highly on operator skill and can be very problematic in comparing results between labs or even between technicians. This project will focus on two areas of high interest to PBR Laboratories, a company that specializes in these comet assays. First, an improved chip design that enhances hydrogel adhesion to the substrate will be produced. Hydrogels tend to have extremely poor adhesion to most surfaces, and chemically treated chips can cost up to several hundred dollars for multi-plate arrays and several dollars for simple treated glass slides. A less expensive option than current technology is highly desirable. Second, a means to produce more uniform hydrogel thicknesses and a system to organize where cells are located on chip with high accuracy will improve uniformity of results. Finally, a means to apply multiple mutagens or doses to cells in a single test will massively increase the throughput of the comet assay for lower costs per test and improved statistical results. This complete system should be as simple as possible so as to lend itself to high automation in future implementations. These project objectives will be accomplished through the use of new plastic manufacturing processes design for microfluidic handling and control which should reduce chip costs while also being compatible with PBR's existing test infrastructure.