High adhesive substrate and microfluidic system for the comet assay

该项目旨在生产一种改进的系统，用于在芯片上运行彗星试验， 控制、均匀性以及比当前解决方案更低的制造成本。彗星试验是一种特殊的 一种基因组测试，用于检测暴露于诱变剂后细胞的遗传损伤。细胞被包裹 在水凝胶中保持它们在适当的位置，暴露于感兴趣的条件下，然后破裂，DNA染色， 荧光可视化施加电场，受损的DNA扩散得更多，提供了一个“彗星”。 像外观一样，而未受损的DNA仍然紧密聚集。虽然彗星试验是非常 有用的，这是一个缓慢的，非常手动的过程，高度依赖于操作员的技能，并可能是非常有问题的， 实验室之间甚至技术人员之间的结果比较。该项目将重点关注两个高领域 PBR实验室是一家专门从事这些彗星分析的公司。首先，改进的芯片设计 其增强了水凝胶对基材的粘附性。水凝胶往往具有极差的 粘附到大多数表面，化学处理芯片可以花费高达数百美元的多板 阵列和几美元的简单处理的载玻片。一个比现有技术更便宜的选择是 非常可取。第二，生产更均匀的水凝胶厚度的手段和组织系统， 其中细胞以高精度位于芯片上将提高结果的均匀性。最后，一种应用方法 在一次试验中对细胞使用多种诱变剂或剂量将大大增加彗星试验的通量 以降低每次测试的成本并改进统计结果。这个完整的系统应该尽可能简单 以便在将来的实施中使其自身具有更高的自动化。这些项目目标将是 通过使用新的塑料制造工艺设计的微流体处理和 控制，这将降低芯片成本，同时也与PBR的现有测试基础设施兼容。

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.