Nanodroplets Based High Throughput Single-Cell PCR System

描述（由申请人提供）：PCR系统细胞内容物的化学分析，如核酸和蛋白质，对生物学，医学和制药界具有重要意义。细胞内物质中异常基因和蛋白的检测为疾病早期诊断提供了重要线索。对越来越小的纯细胞群体进行精确分子分析的愿望已经发展成为微基因组学的新兴领域。不幸的是，由于缺乏确保此类研究可靠性所需的过程控制，研究人员倾向于放弃使用微基因组技术。微基因组技术的主要困难是难以处理微量样品，无法制备和操作单细胞，以及无法具有高通量能力。更重要的是，它是极其劳动密集型的，并且需要昂贵的设备来分离单个细胞并对每个细胞进行扩增，因此每次可以测试的细胞数量非常有限。因此，Maxwell Sensors公司建议开发一种基于纳米滴的高通量单细胞PCR （HTSC PCR）芯片/系统，该芯片/系统将为所需靶细胞的病理、肿瘤发生和其他过程的发展提供前所未有的关键信息。HTSC PCR系统结合了纳米液滴、微流体和荧光反转录（RT） PCR技术，以快速、高通量和低成本的方式进行单细胞（SC） RT PCR。这个高度集成的平台被配置为精确地制备和封装在纳米液滴中的单个细胞，同时在纳升体积中执行数百或数千个单细胞PCR，快速分析芯片上的PCR结果，并为临床诊断或生物医学研究目的的大群体单细胞分析开辟道路。纳米液滴的高保真操作和核酸扩增能力的结合为开发强大的自动化仪器提供了可能。

DESCRIPTION (provided by applicant): PCR System Chemical analysis of cellular contents, such as nucleic acids and proteins, is of significant interest to the biological, medical, and pharmaceutical communities. Detection of abnormal gene and protein in the intracellular materials provide important clues for early disease diagnosis. The desire to perform precise molecular analysis on progressively smaller, pure cell populations has developed into the bourgeoning field of microgenomics. Unfortunately, investigators have tended to forgo the use of microgenomic technologies due to a lack of process control needed to ensure reliability of such studies. Key difficulties in microgenomic technologies are the difficulty in handling a minute amount of sample, inability to prepare and manipulate single cell, and inability to have high throughput capability. More importantly, it is extremely labor intensive and requires expensive equipment to isolate single cells and to perform amplification on each cell, so that the cell number that can be tested each time is very limited. Therefore, Maxwell Sensors Inc. proposes to develop a nanodrop based High Throughput Single Cell PCR (HTSC PCR) chip/system that will provide unprecedented information critical to the development of pathology, oncogenesis, and other processes of a desired target cell. The HTSC PCR system combines nanodroplet, microfluidics, and fluorescent reverse transcription (RT) PCR technologies for performing single cell (SC) RT PCR in a quick, high throughput and cost effective fashion. This highly integrated platform is configured to precisely prepare and encapsulate a single cell in a nanodroplet, simultaneously perform hundreds or thousands of single cell PCR in nanoliter volumes, rapidly analyze the PCR results on chip, and open the road to single cell analyses of large populations for clinical diagnostics or biomedical research purpose. The combination of high fidelity manipulation of nanodroplets and the ability to perform nucleic acid amplification offers the possibility of developing powerful automated instruments.