IMAGING BARRIERS TO MOLECULAR DIFFUSION BY PAIR CORRELATION FUNCTIONS

通过成对相关函数对分子扩散的障碍进行成像

• 批准号: 8365762
• 负责人: ENRICO GRATTON
• 金额: $ 4.27万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-20 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365762
• 关键词: Cells; Complex; Diffusion; Event; Fluorescence; Funding; Grant; Image; Kinetics; Label; Laboratories; Lasers; Life; Location; Measurement; Measures; Modeling; Molecular; National Center for Research Resources; Nuclear Envelope; Nuclear Pore; Principal Investigator; Procedures; Research; Research Infrastructure; Resources; Source; Techniques; Time; Transport Process; United States National Institutes of Health; base; cost; novel; single molecule

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. I study the intracellular diffusivity of a model substrate (NLS-GFP) by using a novel and unconventional fluorescence correlation technique, based on pair correlation functions. I detect molecular transport events by measuring the time a single molecule takes to move from a specific location to another within the cell. Here is the novelty with respect to the traditional FCS approach that measures the time a molecule takes to cross the laser beam. By this approach obstacle to diffusion can be detected both within cellular compartments (restrictions to free diffusion) and across the nuclear envelope barrier (restrictions due to the presence of nuclear pore channels). In both cases the measurement affords a quantitative description of the kinetic steps of NLS-GFP transport at the single molecule level, and in living cells, without the need for complex labeling procedures and cell manipulations.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 我通过基于配对相关函数的新型且非常规荧光相关技术研究模型底物（NLS-GFP）的细胞内扩散率。我通过测量单个分子从特定位置移到细胞内另一个位置的时间来检测分子传输事件。这是关于传统FCS方法的新颖性，该方法测量了分子穿过激光束所需的时间。通过这种方法，可以在细胞室内（自由扩散的限制）和整个核包膜屏障（由于存在核孔通道引起的限制），可以检测到扩散的障碍。在这两种情况下，测量均提供了单个分子水平和活细胞中NLS-GFP传输动力学步骤的定量描述，而无需复杂的标记程序和细胞操作。