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Determination of supramolecular structure and spatial distribution of protein complexes in living cells

机译：活细胞中超分子结构的确定和蛋白质复合物的空间分布

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Resonant energy transfer from an optically excited donor molecule to a non-excited acceptor molecule residing nearby is widely used to detect molecular interactions in living cells. To date, resonant energy transfer has been used to obtain stoichiometric information, such as the number of proteins forming a complex, for a handful of proteins, but only after performing sequential scans of the emission wavelengths, excitation wavelengths, or sometimes both. During this lengthy process of measurement, the molecular makeup of a cellular region may change, limiting the applicability of resonant energy transfer to the determination of cellular averages. Here, we demonstrate a method for the determination of protein complex size, configuration, and spatial distribution in single living cells. It relies on a spectrally resolved two-photon microscope, a simple but competent theory, and a judicious selection of fluorescent tags. This approach eventually may lead to tracking the dynamics of individual molecular complexes inside living cells.
机译：从光学激发的供体分子到附近的非激发受体分子的共振能量转移被广泛用于检测活细胞中的分子相互作用。迄今为止，共振能量转移已被用于获得少量蛋白质的化学计量信息，例如形成复合物的蛋白质数量，但仅在对发射波长，激发波长或有时对两者进行顺序扫描之后才能获得。在漫长的测量过程中，细胞区域的分子组成可能会发生变化，从而将共振能量转移的适用范围限制在确定细胞平均数上。在这里，我们演示了确定单个活细胞中蛋白质复合物的大小，构型和空间分布的方法。它依靠光谱解析的双光子显微镜，简单但有效的理论以及对荧光标签的明智选择。这种方法最终可能导致追踪活细胞内单个分子复合物的动力学。

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