Nanodroplets Based High Throughput Single-Cell PCR System

基于纳米液滴的高通量单细胞 PCR 系统

• 批准号: 7399762
• 负责人: John Collins
• 金额: $ 15万
• 依托单位: MAXWELL SENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2009-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7399762
• 关键词: Actins; Base Ratios; Biological; Biological Assay; Biomedical Engineering; Biomedical Research; California; Cell Count; Cell Separation; Cells; Cellular Structures; Chemicals; Clinical; Collaborations; Communities; Complementary DNA; Computer software; Condition; Cytolysis; Data; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Dyes; Elements; Encapsulated; Ensure; Enzymes; Equipment; Evaluation; Feasibility Studies; Figs - dietary; Fluorescence; Gene Proteins; Genomics; Germ Cells; Goals; HIV; Halogens; Heating; Ifosfamide/Lomustine/Vinblastine; Image; Individual; Lead; Light; Liquid substance; Medical; Messenger RNA; Microfluidics; Microscope; Molecular Analysis; Monitor; Nucleic Acids; Numbers; Oils; Operative Surgical Procedures; Optics; Pathology; Performance; Peripheral Blood Mononuclear Cell; Pharmacologic Substance; Phase; Polymerase Chain Reaction; Population; Procedures; Process; Proteins; Protocols documentation; Pump; Purpose; RNA; RNA-Directed DNA Polymerase; Range; Rate; Reagent; Research Personnel; Reverse Transcription; Sampling; Screening procedure; Sensitivity and Specificity; Signal Transduction; Sorting - Cell Movement; Source; Specificity; Speed; Syringes; System; Technology; Temperature; Testing; Time; Tungsten; Universities; Viral; Width; Work; aqueous; base; cost; cost effective; design; desire; detector; improved; instrument; interest; nanoDroplet; nanolitre; programs; prototype; research study; sensor; simulation; size; success; tumorigenesis; viral RNA

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.

描述（由申请人提供）：PCR 系统 细胞内容物（例如核酸和蛋白质）的化学分析对于生物、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质的检测为早期疾病诊断提供了重要线索。 对越来越小的纯细胞群进行精确分子分析的愿望已经发展成为新兴的微观基因组学领域。 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观基因组技术。 微观基因组技术的主要难点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，它的劳动强度极大，需要昂贵的设备来分离单个细胞并对每个细胞进行扩增，使得每次可以测试的细胞数量非常有限。 因此，Maxwell Sensors Inc. 提议开发一种基于 Nanodrop 的高通量单细胞 PCR (HTSC PCR) 芯片/系统，它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，以快速、高通量和经济高效的方式执行单细胞 (SC) RT PCR。 这个高度集成的平台配置为精确制备单细胞并将其封装在纳米液滴中，同时在纳升体积中进行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的大群体单细胞分析开辟道路。纳米液滴的高保真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。