2000-fold parallelized dual-color STED fluorescence nanoscopy

被引：75

作者：

Bergermann, Fabian ^{[1
]}

Alber, Lucas ^{[1
]}

Sahl, Steffen J. ^{[2
]}

Engelhardt, Johann ^{[1
]}

Hell, Stefan W. ^{[1
,2
]}

机构：

[1] German Canc Res Ctr, D-69120 Heidelberg, Germany

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

来源：

OPTICS EXPRESS | 2015年 / 23卷 / 01期

关键词：

STRUCTURED-ILLUMINATION MICROSCOPY; STIMULATED-EMISSION; RESOLUTION LIMIT; DIFFRACTION; FIELD; DIFFUSION; DEPLETION; BREAKING; BARRIER;

D O I：

10.1364/OE.23.000211

中图分类号：

O43 [光学];

学科分类号：

070207 [光学];

摘要：

Stimulated Emission Depletion (STED) nanoscopy enables multi-color fluorescence imaging at the nanometer scale. Its typical single-point scanning implementation can lead to long acquisition times. In order to unleash the full spatiotemporal resolution potential of STED nanoscopy, parallelized scanning is mandatory. Here we present a dual-color STED nanoscope utilizing two orthogonally crossed standing light waves as a fluorescence switch-off pattern, and providing a resolving power down to 30 nm. We demonstrate the imaging capabilities in a biological context for immunostained vimentin fibers in a circular field of view of 20 mu m diameter at 2000-fold parallelization (i.e. 2000 "intensity minima"). The technical feasibility of massively parallelizing STED without significant compromises in resolution heralds video-rate STED nanoscopy of large fields of view, pending the availability of suitable high-speed detectors. (C) 2015 Optical Society of America

引用

收藏

页码：211 / 223

页数：13

相关论文

共 32 条

[1]

Nanoscopy in a Living Mouse Brain [J].

Berning, Sebastian ;

Willig, Katrin I. ;

Steffens, Heinz ;

Dibaj, Payam ;

Hell, Stefan W. .

SCIENCE, 2012, 335 (6068) :551-551

[2]

Imaging intracellular fluorescent proteins at nanometer resolution [J].

Betzig, Eric ;

Patterson, George H. ;

Sougrat, Rachid ;

Lindwasser, O. Wolf ;

Olenych, Scott ;

Bonifacino, Juan S. ;

Davidson, Michael W. ;

Lippincott-Schwartz, Jennifer ;

Hess, Harald F. .

SCIENCE, 2006, 313 (5793) :1642-1645

[3]

Parallelized STED fluorescence nanoscopy [J].

Bingen, Pit ;

Reuss, Matthias ;

Engelhardt, Johann ;

Hell, Stefan W. .

OPTICS EXPRESS, 2011, 19 (24) :23716-23726

[4]

Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses [J].

Bueckers, Johanna ;

Wildanger, Dominik ;

Vicidomini, Giuseppe ;

Kastrup, Lars ;

Hell, Stefan W. .

OPTICS EXPRESS, 2011, 19 (04) :3130-3143

[5]

Nanoscopy with more than 100,000 'doughnuts' [J].

Chmyrov, Andriy ;

Keller, Jan ;

Grotjohann, Tim ;

Ratz, Michael ;

d'Este, Elisa ;

Jakobs, Stefan ;

Eggeling, Christian ;

Hell, Stefan W. .

NATURE METHODS, 2013, 10 (08) :737-+

[6]

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

[7]

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

[8]

Focal spots of size λ/23 open up far-field florescence microscopy at 33 nm axial resolution -: art. no. 163901 [J].

Dyba, M ;

Hell, SW .

PHYSICAL REVIEW LETTERS, 2002, 88 (16) :4-163901

[9]

Fluorescence nanoscopy by ground-state depletion and single-molecule return [J].

Foelling, Jonas ;

Bossi, Mariano ;

Bock, Hannes ;

Medda, Rebecca ;

Wurm, Christian A. ;

Hein, Birka ;

Jakobs, Stefan ;

Eggeling, Christian ;

Hell, Stefan W. .

NATURE METHODS, 2008, 5 (11) :943-945

[10]

A STED Microscope Designed for Routine Biomedical Applications [J].

Goerlitz, Frederik ;

Hoyer, Patrick ;

Falk, Henning J. ;

Kastrup, Lars ;

Engelhardt, Johann ;

Hell, Stefan W. .

PROGRESS IN ELECTROMAGNETICS RESEARCH-PIER, 2014, 147 :57-68

← 1 2 3 4 →