Microfluidic PCR System for Single Cell Transcriptional Analysis

描述(由申请人提供)： 这项建议的目标是获得一个足够灵敏的PCR系统，用于单细胞转录分析。该系统被称为来自Fluidigm的BioMark HD系统，能够进行实时、终端和数字PCR。该系统的关键方面是在一个芯片上使用集成的微流控通道，允许在纳升体积和皮克量的RNA中同时进行聚合酶链式反应。具体地说，单个细胞(每次最多96次)的转录本(每次最多96次)可以在一次运行中进行检测。这一独特的能力将极大地扩展密歇根大学可以解决的多个调查方向的研究问题。具体的例子包括转移研究，如播散性肿瘤细胞生物学或癌症干细胞生物学。这个应用程序支持14个主要用户，他们总共拥有20个R01，一个P01中有4个项目，两个不同P50孢子中有3个项目，U01财团中有2个项目，总共31个NIH资助项目中，NIH主任奖和U54奖各1个。该小组由多个部门以及医学和牙科学校的成员组成。目前，密歇根大学还没有进行单细胞转录分析的能力，这种设备将使我们能够将研究扩展到这一非常重要的领域。这些设备将被放置在癌症中心大楼的中心位置，以便许多调查人员能够接触到。已经计划使用该设备的项目将利用86.8%的可用设备时间，这表明对该设备的需求很高。研究项目与NIH改善健康的目标直接相关，包括癌症转移、癌症治疗、血管生成、骨生物学和与年龄相关的疾病的研究。该设备将由一名经验丰富的技术人员、一名生物信息学专家和一个内部咨询委员会提供支持。一个吸引和培训新用户的系统已经到位，这样NIH资助的其他调查人员将从这些设备中受益。头三年承诺为维护和技术费用提供大量的机构支助，以确保该系统的长期可行性(三年共计100500美元)。将建立以用户为基础的充值系统，以继续长期使用。此外，还有为设备提供空间的机构承诺。总而言之，使用这一尖端成像系统将大大加快密歇根大学大量由NIH资助的研究人员的研究步伐。

DESCRIPTION (provided by applicant): The objective of this proposal is to acquire a PCR system that is sensitive enough for single cell transcriptional analysis. The system, called the Biomark HD System from Fluidigm is capable of real- time, end-point and digital PCR. The key aspects of this system are the use of integrated microfluidic channels on one chip that allow for simultaneous PCR reactions in nanoliter volumes and picogram amounts of RNA. In concrete terms, transcripts (up to 96 per run) from individual cells (up to 96 per run) can be assayed in a single run. This unique capability will greatly expand the research questions that can be addressed at University of Michigan for multiple investigative directions. Specific examples include metastasis studies such as disseminated tumor cell biology or cancer stem cell biology. This application supports 14 major users who together have 20 R01s, 4 projects in one P01, 3 projects in two different P50 SPORES, 2 projects in U01 consortia and 1 each of a NIH Director's Award and a U54 award for a total of 31 NIH-funded projects. The group consists of members of multiple Departments and the Medical and Dental Schools. Currently, there is not capability to perform single cell transcriptional analysis at University of Michigan and this equipment will enable us to expand studies into this very important area. The equipment will be placed in a central location in the Cancer Center Building so that it is accessible to many investigators. The projects already planning to use this equipment will utilize 86.8% of the available equipment time indicating the high demand for this equipment. Research projects are directly related to NIH goals of improving health and include studies on cancer metastasis, cancer therapeutics, angiogenesis, bone biology and age- related disease. The equipment will be supported by an experienced technical staff, a bioinformatics expert and an internal advisory committee. A system to attract and train new users is in place so that other NIH-funded investigators will benefit from the equipment. Significant institutional support for both maintenance and technical costs has been committed for the initial three years to ensure long- term viability of the system (Total of $100,500 over 3 years). A user-based recharge system will be put into place to continue its long-term use. Additionally, there is institutional commitment of space for the equipment. In summary access to this cutting edge imaging system will greatly accelerate the pace of research at University of Michigan for a large number of NIH-funded investigators.