NEW ENERGY TRANSFER PROBES FOR SINGLE CELL IMAGING

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。自赠款期开始以来，蛋白质与人工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将大分子有效摄取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，膜平面中的横向运动和单个分子的内化，所有测量均不受系综平均的影响。实时单分子成像是能够记录分子运动的电影并跟踪单个大分子或其复合物的空间路径和化学反应的能力。基于罗丹明的荧光盒正在Kevin Burgess的研究小组中合成，并将在我们的实验室中使用单分子光谱工具进行研究。这些盒被证明采用大的斯托克斯位移，使它们适合于在溶液或单细胞中的单分子实验的潜在应用中使用，其中弱荧光必须与散射的激发光和其他背景源区分开。其他可能的应用包括它们用于使用共聚焦显微镜设置的生物样品的双光子成像。

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.