STED microscopy with continuous wave beams

被引：387

作者：

Willig, Katrin I. ^{[1
]}

Harke, Benjamin ^{[1
]}

Medda, Rebecca ^{[1
]}

Hell, Stefan W. ^{[1
]}

机构：

[1] Max Planck Inst Biophys Chem, Dept NanoBiophoton, D-37077 Gottingen, Germany

来源：

NATURE METHODS | 2007年 / 4卷 / 11期

关键词：

D O I：

10.1038/NMETH1108

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report stimulated emission depletion (STED) fluorescence microscopy with continuous wave (CW) laser beams. Lateral fluorescence confinement from the scanning focal spot delivered a resolution of 29 - 60 nm in the focal plane, corresponding to a 5 - 8- fold improvement over the diffraction barrier. Axial spot confinement increased the axial resolution by 3.5-fold. We observed three-dimensional (3D) subdiffraction resolution in 3D image stacks. Viable for fluorophores with low triplet yield, the use of CW light sources greatly simplifies the implementation of this concept of far-field fluorescence nanoscopy.

引用

页码：915 / 918

页数：4

共 13 条

[1]

Abbe E., 1873, ARCH MIKROSK ANAT, V9, P413, DOI [10.1007/BF02956173, DOI 10.1007/BF02956173]

[2] Major signal increase in fluorescence microscopy through dark-state relaxation [J].

Donnert, Gerald ;

Eggeling, Christian ;

Hell, Stefan W. .

NATURE METHODS, 2007, 4 (01) :81-86

[3] Macromolecular-scale resolution in biological fluorescence microscopy [J].

Donnert, Gerald ;

Keller, Jan ;

Medda, Rebecca ;

Andrei, M. Alexandra ;

Rizzoli, Silvio O. ;

Luehrmann, Reinhard ;

Jahn, Reinhard ;

Eggeling, Christian ;

Hell, Stefan W. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (31) :11440-11445

[4] BREAKING THE DIFFRACTION RESOLUTION LIMIT BY STIMULATED-EMISSION - STIMULATED-EMISSION-DEPLETION FLUORESCENCE MICROSCOPY [J].

HELL, SW ;

WICHMANN, J .

OPTICS LETTERS, 1994, 19 (11) :780-782

[5] Highly nonlinear photodamage in two-photon fluorescence microscopy [J].

Hopt, A ;

Neher, E .

BIOPHYSICAL JOURNAL, 2001, 80 (04) :2029-2036

[6] Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission [J].

Klar, TA ;

Jakobs, S ;

Dyba, M ;

Egner, A ;

Hell, SW .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (15) :8206-8210

[7]

LAKOWICZ JR, 1983, PRINCIPLES FLUROSCEN

[8]

PAWLEY JB, 2006, HDB BIOL CONFOCAL MI

[9] Anatomy and dynamics of a supramolecular membrane protein cluster [J].

Sieber, Jochen J. ;

Willig, Katrin I. ;

Kutzner, Carsten ;

Gerding-Reimers, Claas ;

Harke, Benjamin ;

Donnert, Gerald ;

Rammner, Burkhard ;

Eggeling, Christian ;

Hell, Stefan W. ;

Grubmueller, Helmut ;

Lang, Thorsten .

SCIENCE, 2007, 317 (5841) :1072-1076

[10] Nanoscale resolution in the focal plane of an optical microscope [J].

Westphal, V ;

Hell, SW .

PHYSICAL REVIEW LETTERS, 2005, 94 (14)

← 1 2 →