Microdevice for Direct DNA Purification

描述(申请人提供)：随着人们对组织切片中细胞的分子分类的兴趣与日俱增，快速有效地从所选细胞中提取DNA的能力将是至关重要的。该项目的总体目标是开发一种基于微芯片的样本制备方法，用于从组织样本中高效、低成本地提取DNA-这种微型设备可以轻松地容纳血液或其他细胞来源。这种微型设备将使用最先进的微制造技术，结合流体控制的芯片上细胞裂解和固相萃取化学来制造。该项目结合了HTMicro用于轻松制造复杂、高表面积微结构的工业能力，以及弗吉尼亚大学开发的能够实现高效和高容量DNA提取的表面修饰化学。微型设备将使用各种类型和质量不同的样本进行测试，提取效率将使用实时聚合酶链式反应来确定。作为与当前激光捕获显微切割仪器集成的展示，该微设备将被制造成直接接受来自Arcturus PixCell IIe的盖子，该盖子将在其底部表面包含绑定到乙烯-乙酸乙烯酯聚合物膜上的选定细胞。组织样本将被收集，并由我们在UVA的外科病理学合作者进行激光显微解剖-正常和恶性细胞的样本将被分析。最终的设备将提供微设备的快速分析、高提取效率和高通量优势，此外，预计将提供比当前传统或微芯片技术更高的容量和更低的每设备成本。

DESCRIPTION (provided by applicant): With an ever-increasing interest in the molecular typing of cells from histologic tissue sections, the ability rapidly and efficiently extract DNA from the selected cells will be of paramount importance. The overall goal of this project is to develop a microchip-based sample preparation method for high efficiency, low-cost extraction of DNA from tissue samples - this microdevice will easily accommodate blood or other cells sources. The microdevices will be created using state-of-the-art microfabrication techniques coupled with fluidically controlled on-chip cell lysis and solid phase extraction chemistries. This project couples the industrial capabilities of HT Micro for facile fabrication of complex, high surface area microstructures, with surface modification chemistries developed at the University of Virginia that enable efficient and high capacity DNA extraction. The microdevices will be tested using a variety of sample varying in type and quality, and extraction efficiency will be determined using real-time PCR. As a demonstration of integration with current laser-capture microdissection instruments, the microdevice will be fabricated to directly accept the cap from the Arcturus Pixcell IIe which will contain the selected cells bound to an ethylene vinyl acetate polymer membrane on its bottom surface. The tissue samples will be collected and laser microdissected by our surgical pathology collaborator here at UVa - samples of normal and malignant cells will be analyzed. The final device will offer the rapid analysis, high extraction efficiency, and high-throughput advantages of microdevices and, in addition, is expected to offer higher capacity, and lower cost-per-device than current conventional or microchip techniques.