G. Seisenberger et al., Real-time single-molecule imaging of the infection pathway of an adeno-associated virus, SCIENCE, 294(5548), 2001, pp. 1929-1932
We describe a method, based on single-molecule imaging, that allows the rea
l-time visualization of the infection pathway of single viruses in living c
ells, 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-associat
ed viruses (AAV) during their infection pathway into living HeLa cells: (i)
consecutive virus touching at the cell surface and fast endocytosis; (il)
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 cytopl
asm and in nuclear tubular structures. The real-time visualization of the i
nfection pathway of single AAVs shows a much faster infection than was gene
rally observed so far.