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

在本CAREER项目中，PI将研究IgE受体系统的预聚簇。所提出的方法是使用荧光探针的间歇性和独立的波动，所述荧光探针是光可切换的或自然间歇的，以提取个体轨迹，即使当存在具有低于衍射极限的尺寸的荧光团簇时。该方法是基于隐马尔可夫模型中的参数和状态，这种方法将进一步发展。该项目的具体目标是（1）使用可光开关或自然间歇的荧光探针来寻找高密度膜组分的单膜蛋白轨迹;（2）使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景观;（3）利用分子间的相互作用来确定细胞膜的能量分布。（3）利用所建立的技术，对RBL-2 H3细胞IgE结合受体的行为进行了研究。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有偿研究指导。与荷兰德尔夫特技术大学定量成像组的学生交换计划将允许研究生或有前途的本科生每年进行长达3个月的国际学习。将根据PI部门的研究兴趣开发高级生物物理学课程。将邀请美国国内的演讲者在生物物理学系列研讨会上就相关研究发表演讲。

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.