4Pi-confocal microscopy of live cells

被引:52
作者
Bahlmann, K [1 ]
Jakobs, S [1 ]
Hell, SW [1 ]
机构
[1] Max Planck Inst Biophys Chem, High Resolut Opt Microscopy Grp, D-37070 Gottingen, Germany
关键词
resolution; 4Pi; confocal; E; coli;
D O I
10.1016/S0304-3991(00)00092-9
中图分类号
TH742 [显微镜];
学科分类号
摘要
By coherently adding the spherical wavefronts of two opposing lenses, two-photon excitation 4Pi-confocal fluorescence microscopy has achieved three-dimensional imaging with an axial resolution 3-7 times better than confocal microscopy. So far this improvement was possible only in glycerol-mounted, fixed cells. Here we report 4Pi-confocal microscopy of watery objects and its application to the imaging of live cells. Water immersion of 4Pi-confocal microscopy of membrane stained live Escherichia coli bacteria attains a 4.3-fold better axial resolution as compared to the best water immersion confocal microscope. The resolution enhancement results into a vastly improved three-dimensional representation of the bacteria. The first images of live biological samples with an all-directional resolution in the 190-280 nm range are presented here, thus establishing a new resolution benchmark in live-cell microscopy. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:155 / 164
页数:10
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