APPLICATIONS OF HIGH SENSITIVITY DETECTION

高灵敏度检测的应用

• 批准号: 7598381
• 负责人: BABETTA L MARRONE
• 金额: $ 12.58万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598381
• 关键词: Area; Biochemical; Biological; Categories; Class; Color; Computer Retrieval of Information on Scientific Projects Database; DNA; Detection; Development; Fingerprint; Flow Cytometry; Funding; Genome; Genomics; Goals; Grant; Individual; Institution; Measurement; Measures; Methods; Mitochondria; Phase; Population Heterogeneity; Research; Research Personnel; Resources; Scanning; Science; Source; United States National Institutes of Health; base; falls; instrument; instrumentation; light scattering; molecular assembly/self assembly; particle; research and development; single molecule; size

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The overall goal of this project is to expand the biological applications of High Sensitivity Flow Cytometry (HSFCM) through methods developed and through instrumentation improvements leading to a multiparameter measurement capability. Flow cytometry has enabled major advances in the biomedical sciences by providing rapid, quantitative and sensitive multiparameter measurements of individual cellular particles. Individual analysis produces information on population heterogeneity that is not revealed by ensemble analysis and allows more precise measurement of individual attributes than is possible when measurement is done in bulk phase. However, one limitation of conventional flow cytometry is the inability to measure small particles less than 0.5 ¿m or dim particles having less than several hundred fluorescent molecules. A wide variety of important biological particles, molecules, and molecular assemblies fall into these categories. Our approach is to access this measurement domain through enhanced capability HSFCM. We have identified 3 important biological areas to focus on: bacterial identification by genome analysis; DNA single molecule scanning applications in high throughput genomics; and mitochondrial genomic and biochemical analysis. Specific instrument enhancements in multi-color and light scatter detection will enable these new applications. The research plan is divided into four related Specific Aims: DNA Fragment Size-Based Analysis of Bacterial Fingerprints; High Throughput Scanning of Single DNA Molecules; Analysis of Individual Mitochondria; and High Sensitivity Instrumentation Development. The results of this research and development will create a new class of high sensitivity flow cytometry instrumentation and will stimulate a new field of flow cytometry at the sub-cellular single molecule level.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 该项目的总体目标是通过开发方法和仪器改进来扩展高灵敏度流式细胞术 (HSFCM) 的生物应用，从而实现多参数测量能力。 流式细胞术通过对单个细胞颗粒提供快速、定量和灵敏的多参数测量，使生物医学科学取得了重大进展。 个体分析产生关于群体异质性的信息，这些信息未通过整体分析揭示，并且允许比在批量阶段进行测量时更精确地测量个体属性。 然而，传统流式细胞术的一个限制是无法测量小于 0.5 µm 的小颗粒或具有少于数百个荧光分子的暗颗粒。 许多重要的生物颗粒、分子和分子组装体都属于这些类别。 我们的方法是通过增强功能的 HSFCM 访问此测量域。 我们确定了需要重点关注的 3 个重要的生物学领域：通过基因组分析进行细菌鉴定； DNA单分子扫描在高通量基因组学中的应用；以及线粒体基因组和生化分析。 多色和光散射检测方面的特定仪器增强功能将使这些新应用成为可能。 该研究计划分为四个相关的具体目标：基于 DNA 片段大小的细菌指纹分析；单 DNA 分子的高通量扫描；个体线粒体分析；和高灵敏度仪器开发。 这项研发成果将创造出新型高灵敏度流式细胞仪仪器，并将在亚细胞单分子水平上开辟流式细胞仪的新领域。