Nanodroplets Based High Throughput Single-Cell PCR System

基于纳米液滴的高通量单细胞 PCR 系统

基本信息

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
项目类别：
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

项目摘要

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系统对细胞含量（例如核酸和蛋白质）的化学分析，对生物，医学和药物群落具有重大感兴趣。在细胞内材料中检测异常基因和蛋白质为早期疾病诊断提供了重要的线索。对逐渐较小的纯细胞种群进行精确分子分析的愿望已发展为微基组学的资产阶级领域。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。微基因组技术的关键困难是处理一定量的样本，无法制备和操纵单细胞以及无法具有较高吞吐量能力的困难。更重要的是，它是极度强度的，需要昂贵的设备来分离单个单元格并在每个单元格上进行放大，以便每次可以测试的单元格非常有限。因此，Maxwell Sensors Inc.提议开发基于纳米体的高吞吐量单细胞PCR（HTSC PCR）芯片/系统，该芯片/系统将提供前所未有的信息，对病理学，肿瘤发生和所需靶细胞的其他过程至关重要。 HTSC PCR系统结合了纳米光，微流体和荧光逆转录（RT）PCR技术，用于以快速，高吞吐量和成本效益的方式执行单细胞（SC）RT PCR。这个高度集成的平台配置为精确准备和封装在纳米光中的单个单元，同时在纳米素体积中同时执行数百或数千个单细胞PCR，在芯片上快速分析PCR结果，并为大型群体分析的临床诊断或生物医学研究目的的单细胞分析开放。纳米光的高保真操作与执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。