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测试，具有更高的 控制、一致性以及比当前解决方案更低的制造成本。彗星测试是一种特殊的 一种基因组测试，寻找暴露于诱变剂后对细胞造成的基因损伤。单元格被封装 在水凝胶中保持它们的位置，暴露在感兴趣的条件下，然后破裂和DNA染色 荧光可视化。施加电场后，受损的DNA传播得更广，形成了一颗“彗星” 外观相似，而未受损的DNA保持紧密聚集。虽然彗星测试是非常 很有用，这是一个缓慢的、非常手动的过程，高度依赖操作员技能，并且在 在实验室之间，甚至在技术人员之间比较结果。该项目将集中在两个领域的高 感兴趣的是PBR实验室，一家专门从事这些彗星分析的公司。第一，改进了芯片设计 将产生增强水凝胶与基材粘附性的材料。水凝胶往往具有极差的 粘合到大多数表面和化学处理的芯片上，多层板的成本可能高达数百美元 阵列和几美元的简单处理的玻璃幻灯片。一个比当前技术更便宜的选择是 非常可取。第二，生产更均匀水凝胶厚度的手段和组织系统 芯片上单元位置的高精度将提高结果的一致性。最后，一种适用于 一次测试中对细胞的多个诱变剂或剂量将极大地增加彗星试验的吞吐量 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 以便在未来的实现中实现高度自动化。这些项目目标将是 通过使用新的塑料制造工艺设计用于微流控处理和 控制，这将降低芯片成本，同时也兼容PBR现有的测试基础设施。

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.