Nanodroplets Based High Throughput Single-Cell PCR System

描述（由申请人提供）：PCR系统细胞内容物（如核酸和蛋白质）的化学分析对生物、医学和制药界具有重大意义。细胞内异常基因和蛋白质的检测为疾病的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观基因组学的新兴领域。 不幸的是，调查人员倾向于放弃使用 微基因组技术，由于缺乏过程控制，需要确保这种研究的可靠性。 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，它劳动强度极大，需要昂贵的设备来分离单个细胞并对每个细胞进行扩增，因此每次可以检测的细胞数量非常有限。 因此，麦克斯韦传感器公司 提出开发一种基于纳米滴的高通量单细胞PCR（HTSC PCR）芯片/系统，其将提供对所需靶细胞的病理学、肿瘤发生和其他过程的发展至关重要的前所未有的信息。 HTSC PCR系统结合了纳米液滴、微流体和荧光逆转录（RT）PCR技术，用于以快速、高通量和具有成本效益的方式进行单细胞（SC）RT PCR。 这种高度集成的平台被配置为精确地制备和封装纳米液滴中的单细胞，同时进行纳升体积的数百或数千个单细胞PCR，在芯片上快速分析PCR结果，并为临床诊断或生物医学研究目的打开大群体单细胞分析的道路。纳米液滴的高保真度操作和进行核酸扩增的能力的结合提供了开发强大的自动化仪器的可能性。

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.