Microdevice for Direct DNA Purification

描述（由申请人提供）：随着对组织学组织切片细胞分子分型的兴趣不断增加，从选定细胞中快速有效地提取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品制备方法，用于高效率、低成本地从组织样本中提取DNA——这种微型设备将很容易适应血液或其他细胞来源。微型器件将采用最先进的微制造技术，结合流体控制芯片上的细胞裂解和固相萃取化学技术。该项目将HT Micro的工业能力与弗吉尼亚大学开发的表面修饰化学技术结合起来，可以轻松制造复杂、高表面积的微结构，从而实现高效、高容量的DNA提取。将使用各种不同类型和质量的样品对微设备进行测试，并使用实时PCR确定提取效率。作为与当前激光捕获显微解剖仪器集成的演示，该微设备将被制造成直接接受来自Arcturus Pixcell IIe的帽，该帽将包含选定的细胞，并将其与底部表面的乙烯醋酸乙烯聚合物膜结合。组织样本将被收集并由我们在弗吉尼亚大学的外科病理学合作者进行激光显微解剖，正常细胞和恶性细胞的样本将被分析。最终的设备将提供快速分析、高萃取效率和微设备的高通量优势，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台设备成本。

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.