通过受激发射打破衍射分辨率屏障的荧光显微镜。
Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission.
作者信息
Klar T A, Jakobs S, Dyba M, Egner A, Hell S W
机构信息
Max-Planck-Institute for Biophysical Chemistry, High Resolution Optical Microscopy Group, 37070 Göttingen, Germany.
出版信息
Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8206-10. doi: 10.1073/pnas.97.15.8206.
Abstract
The diffraction barrier responsible for a finite focal spot size and limited resolution in far-field fluorescence microscopy has been fundamentally broken. This is accomplished by quenching excited organic molecules at the rim of the focal spot through stimulated emission. Along the optic axis, the spot size was reduced by up to 6 times beyond the diffraction barrier. The simultaneous 2-fold improvement in the radial direction rendered a nearly spherical fluorescence spot with a diameter of 90-110 nm. The spot volume of down to 0.67 attoliters is 18 times smaller than that of confocal microscopy, thus making our results also relevant to three-dimensional photochemistry and single molecule spectroscopy. Images of live cells reveal greater details.
摘要
远场荧光显微镜中造成有限焦斑尺寸和有限分辨率的衍射障碍已从根本上被打破。这是通过受激发射淬灭焦斑边缘的激发有机分子来实现的。沿光轴方向，光斑尺寸在突破衍射障碍后减小了多达6倍。在径向方向上同时实现了2倍的改善，形成了直径为90 - 110纳米的近乎球形的荧光光斑。低至0.67飞升的光斑体积比共聚焦显微镜的光斑体积小18倍，因此我们的结果也与三维光化学和单分子光谱学相关。活细胞图像显示出更多细节。
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