CAREER: A Computational and Analytical Approach to Single-Molecule Fluorescence Imaging for the Quantitative Analysis of Protein Interactions in Living Cells.

职业：用于定量分析活细胞中蛋白质相互作用的单分子荧光成像计算和分析方法。

• 批准号: 0954836
• 负责人: Keith Lidke
• 金额: $ 51.23万
• 依托单位: University of New Mexico
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-10-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0954836&HistoricalAwards=false
• 关键词: CAREER; Computational; Analytical; Approach; Single

In this CAREER project the PI will study pre-clustering of the IgE receptor system. The proposed approach is to use the intermittent and independent fluctuations of fluorescent probes that are either photo-switchable or naturally intermittent to extract individual trajectories even when there are clusters of fluorophores with sizes below the diffraction limit. The approach is based on finding parameters and states in hidden Markov models and this approach will be further developed. The specific aims of this project are (1) to use fluorescent probes that are either photo-switchable or naturally intermittent to find single membrane protein trajectories of high density membrane components; (2) to use the trajectories of closely spaced proteins in conjunction with physical models to extract the binding kinetics between single molecules and to map out the energy landscape of the cell membrane; (3) using the developed techniques, to characterize the behavior of the IgE binding receptor of RBL-2H3 cells. The project will provide multidisciplinary training for a graduate student as well as undergraduate students and offer annual, paid research mentorships to high school students in the Albuquerque, New Mexico area. A student exchange program with the Quantitative Imaging group at Delft Technical University, Netherlands will allow up to 3 months annual international study for graduate students or promising undergraduates. An upper level Biophysics course will be developed based on the research interests of the PI's department. Speakers will be invited from within the U.S. to give talks on related research in a biophysics seminar series.

在这个职业项目中，PI 将研究 IgE 受体系统的预聚类。所提出的方法是利用荧光探针的间歇性和独立波动来提取单个轨迹，即使存在尺寸低于衍射极限的荧光团簇，这些荧光探针可以光切换或自然间歇性地提取单个轨迹。该方法基于寻找隐马尔可夫模型中的参数和状态，并且该方法将得到进一步发展。该项目的具体目标是（1）使用光可切换或自然间歇的荧光探针来寻找高密度膜成分的单膜蛋白轨迹； （2）利用紧密间隔的蛋白质的轨迹结合物理模型来提取单分子之间的结合动力学并绘制细胞膜的能量图谱； (3)使用开发的技术，表征RBL-2H3细胞的IgE结合受体的行为。该项目将为研究生和本科生提供多学科培训，并为新墨西哥州阿尔伯克基地区的高中生提供年度付费研究指导。与荷兰代尔夫特理工大学定量成像小组的学生交换计划将为研究生或有前途的本科生提供长达 3 个月的年度国际学习机会。将根据 PI 部门的研究兴趣开发高级生物物理学课程。将邀请美国境内的演讲者在生物物理学研讨会系列中就相关研究进行演讲。