NEW ENERGY TRANSFER PROBES FOR SINGLE CELL IMAGING

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。自资助期开始以来，蛋白质与人工细胞膜和活细胞膜相互作用的单分子研究取得了显着进展。蛋白质与细胞膜的相互作用对于有效地将大分子吸收到活细胞中是必不可少的。单分子荧光方法使我们能够研究主要事件，如大分子与膜的结合，膜平面内的横向运动和单个分子的内化，所有的测量都不需要系综平均。实时单分子成像是记录分子运动的电影，并跟踪单个大分子或其络合物的空间路径和化学反应的能力。Kevin Burgess的研究小组正在合成基于罗丹明的荧光盒，并将在我们的实验室使用单分子光谱工具进行研究。这些盒使用了大的斯托克斯位移，使它们适合于在溶液中或在单池中进行单分子实验的潜在应用，其中必须将微弱的荧光与散射激发光和其他背景源区分开来。其他可能的应用包括使用共聚焦显微镜装置对生物样品进行双光子成像。

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. Single molecule studies of protein interactions with artificial and living cell membranes have progressed significantly since the commencement of the grant period. Interactions of proteins with the cell membrane are essential for efficient uptake of macromolecules into living cells. Single molecule fluorescence approaches have allowed us to study primary events such as the binding of macromolecules to membranes, lateral motion in the membrane plane and internalization for individual molecules, with all measurements free from ensemble averaging. Real-time single molecule imaging is the ability to record movies of molecular motions and to follow the spatial pathways and chemical reactions of individual macromolecules or their complexes. Rhodamine based fluorescent cassettes are being synthesized in the research group of Kevin Burgess and will be investigated in our laboratories using single molecule spectroscopic tools. These cassettes were shown to employ large stokes shifts making them suitable to be used in potential applications of single molecule experiments in solution or in single cells, where the weak fluorescence has to be distinguished from scattered excitation light and other background sources. Other possible applications include their use for two-photon imaging of biological samples using a confocal microscope setup.