PHONE CONSULTATION: CONFOCAL MICROSCOPY

在DAMOP会议上的讲话，圣达菲1998年5月27日至30日会议I1- 分子生物学的光学技术。NVITED会议，周五 5月29日上午，FCS，1972年引入的一种技术(Magde等人 1972年，Appl.太棒了。让我们来吧。29：705)，目前受雇于 分子扩散和聚集的超灵敏研究， 光物理动力学和快速化学反应。在以下位置执行时 单光子或双光子激发的共焦几何(Denk等人) 1990，Science 248：73)光稳定性染料，它的灵敏度使我们能够 要遵循稀疏生物相关因子的动态，例如 溶液和活细胞中的药物和病原体。最低限度 干扰观察系统的生物功能，FCS 使我们能够解决分子中的各种重要问题 生物学。我们报道了绿色荧光蛋白(GFP)，一种流行的 体内研究的工具，不仅非常适合细胞内 功能界别，但事实证明是这项研究的完美候选者 分子内动力学。它在100秒内呈现出快速闪烁 ？我们已经确定的S时间范围是由于可逆原型 发色团之国。由NIH-P41.RR04224、NSF-D1R支持 8800278

Talk at DAMOP meeting, Santa Fe May 27-30 1998 Session I1 - Optical Techniques in Molecular Biology. NVITED session, Friday morning, May 29 FCS, a technique introduced in 1972 (Magde et al. 1972, Appl. Phys. Lett. 29:705), is currently being employed for ultrasensitive studies of molecular diffusion and aggregation, photophysical dynamics and fast chemical reactions. When performed in confocal geometries with one- or two-photon excitation (Denk et al. 1990, Science 248:73) of photostable dyes, its sensitivity enables us to follow the dynamics of sparse biologically relevant agents such as drugs and pathogens in solution as well as in living cells. Minimally interfering with biological functions of the observed system, FCS allows us to address a variety of important questions in molecular biology. We report that green fluorescent protein (GFP), a popular tool for in-vivo studies, is not only well suited for intracellular FCS, but moreover turned out to be a perfect candidate for the study of intramolecular dynamics. It exhibits a rapid flickering in the 100 ?s time range that we have identified to be due to reversible proto nation of the chromophore. Supported by NIH-P41.RR04224, NSF-D1R 8800278