Quantitative Aspects of Single Molecule Detection

单分子检测的定量方面

• 批准号: 6865191
• 负责人: RAIMUND J OBER
• 金额: $ 26.16万
• 依托单位: UNIVERSITY OF TEXAS DALLAS
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6865191
• 关键词: analytical method; bioimaging /biomedical imaging; computer program /software; computer system design /evaluation; fluorescence microscopy; image processing; method development; molecular /cellular imaging; nanotechnology

DESCRIPTION (provided by applicant): The advent of technology that allows the detection of single molecules in a cellular environment holds the promise that significant new insights can be gained into molecular processes that occur in a cell. While the promise is great so are the technical problems. This is primarily because the signal emitted by a single molecule is low in comparison to the significant noise sources in the sample and in the detection system. Therefore any quantitative analysis of single molecule experiments is challenging and requires carefully developed tools. The overall goal of this proposal is to add to the available methods for the analysis of single molecule experiments. In particular, we address fundamental problems using methods that are novel to single molecule microscopy but have been successfully used in other engineering disciplines. We will analyze performance limits of single molecule microscopy from a modern point of view. Our specific aims are: 1.) To develop novel analytical tools to investigate the accuracy with which single molecules can be localized. 2.) To investigate existing algorithms and to develop novel algorithms for the localization of single molecules. 3.) To develop novel resolution criteria for single molecule detection and algorithms for the determination of the distance between two single molecules. 4.) To develop software modules for our existing Matlab based microscope image processing toolbox. We will use approaches based on the Fisher information matrix to obtain novel expressions for the accuracy with which a single molecule can be localized and for the accuracy with which the distance between two single molecules can be measured. Of particular importance is the determination to what extent experimental conditions such as cellular background and noise in the detection system deteriorate the localization and resolution accuracy. Existing algorithms will be evaluated to establish how close their performance is to the performance limit. Novel algorithms will be designed to meet the performance measures. A further criterion will be to assess the sensitivity of the algorithms to experimental artifacts such as outliers in the data. All proposed methods will be tested on immunological systems.

描述（由申请人提供）：允许检测细胞环境中的单个分子的技术的出现有望获得对细胞中发生的分子过程的重要新见解。虽然前景很好，但技术问题也很大。这主要是因为与样品和检测系统中的显著噪声源相比，由单个分子发射的信号较低。因此，单分子实验的任何定量分析都是具有挑战性的，需要精心开发的工具。该提案的总体目标是增加用于分析单分子实验的可用方法。特别是，我们使用的方法是新颖的单分子显微镜，但已成功地用于其他工程学科的基本问题。我们将从现代的角度分析单分子显微镜的性能极限。 我们的具体目标是： 1.）的人。开发新的分析工具，以研究单分子定位的准确性。 2.）的情况。研究现有算法并开发用于单分子定位的新型算法。 3.）第三章开发用于单分子检测的新分辨率标准和用于确定两个单分子之间距离的算法。 4.）为我们现有的基于Matlab的显微镜图像处理工具箱开发软件模块。 我们将使用基于Fisher信息矩阵的方法，以获得新的表达式的准确性，其中一个单一的分子可以被本地化和准确性，可以测量两个单一的分子之间的距离。特别重要的是确定在何种程度上的实验条件，如细胞背景和检测系统中的噪声恶化的定位和分辨率精度。将评估现有算法，以确定其性能与性能极限的接近程度。新的算法将被设计为满足性能指标。另一个标准是评估算法对实验伪影（如数据中的离群值）的敏感性。所有提出的方法都将在免疫系统上进行测试。