Stimulated emission depletion (STED) nanoscopy of a fluorescent protein-labeled organelle inside a living cell

被引：338

作者：

Hein, Birka ^{[1
]}

Willig, Katrin I. ^{[1
]}

Hell, Stefan W. ^{[1
]}

机构：

[1] Max Planck Inst Biophys Chem, Dept Nanobiophoton, D-37070 Gottingen, Germany

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2008年 / 105卷 / 38期

关键词：

fluorescence; GFP; microscopy; resolution;

D O I：

10.1073/pnas.0807705105

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

We demonstrate far-field optical imaging with subdiffraction resolution of the endoplasmic reticulum (ER) in the interior of a living mammalian cell. The diffraction barrier is overcome by applying stimulated emission depletion (STED) on a yellow fluorescent protein tag. Imaging individual structural elements of the ER revealed a focal plane (x, y) resolution of < 50 nm inside the living cell, corresponding to a 4-fold improvement over that of a confocal microscope and a 16-fold reduction in the focal-spot cross-sectional area. A similar gain in resolution is realized with both pulsed- and continuous-wave laser illumination. images of highly convoluted parts of the ER reveal a similar resolution improvement in 3D optical sectioning by a factor of 3 along the optic axis (z). Time-lapse STED recordings document morphological changes of the ER over time. Thus, nanoscale 313 imaging of organelles in the interior of living cells greatly expands the scope of light microscopy in cell biology.

引用

页码：14271 / 14276

页数：6

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