Seisenberger G, Ried M U, Endress T, Büning H, Hallek M, Bräuchle C
Department Chemie, Ludwig-Maximilians-Universität München, Butenandtstrasse 11, D-81377 München, Germany.
Science. 2001 Nov 30;294(5548):1929-32. doi: 10.1126/science.1064103.
We describe a method, based on single-molecule imaging, that allows the real-time visualization of the infection pathway of single viruses in living cells, each labeled with only one fluorescent dye molecule. The tracking of single viruses removes ensemble averaging. Diffusion trajectories with high spatial and time resolution show various modes of motion of adeno-associated viruses (AAV) during their infection pathway into living HeLa cells: (i) consecutive virus touching at the cell surface and fast endocytosis; (ii) free and anomalous diffusion of the endosome and the virus in the cytoplasm and the nucleus; and (iii) directed motion by motor proteins in the cytoplasm and in nuclear tubular structures. The real-time visualization of the infection pathway of single AAVs shows a much faster infection than was generally observed so far.
我们描述了一种基于单分子成像的方法，该方法能够实时可视化单个病毒在活细胞中的感染途径，每个病毒仅用一个荧光染料分子进行标记。对单个病毒的追踪消除了总体平均效应。具有高空间和时间分辨率的扩散轨迹显示了腺相关病毒（AAV）在感染活的HeLa细胞过程中的各种运动模式：（i）病毒在细胞表面连续接触并快速内吞；（ii）内体和病毒在细胞质和细胞核中自由且异常扩散；（iii）在细胞质和核管状结构中由运动蛋白介导的定向运动。单个AAV感染途径的实时可视化显示其感染速度比迄今为止普遍观察到的要快得多。
