Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT

被引：62

作者：

Hoyer, Patrick ^{[1
,2
,3
]}

de Medeiros, Gustavo ^{[3
]}

Balazs, Balint ^{[3
,4
]}

Norlin, Nils ^{[3
]}

Besir, Christina ^{[3
]}

Hanne, Janina ^{[1
,2
,5
]}

Kraeusslich, Hans-Georg ^{[5
]}

Engelhardt, Johann ^{[1
,2
]}

Sahl, Steffen J. ^{[6
]}

Hell, Stefan W. ^{[1
,2
,6
]}

Hufnagel, Lars ^{[3
]}

机构：

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

[2] Bioquant, D-69120 Heidelberg, Germany

[3] European Mol Biol Lab, Cell Biol & Biophys Unit, D-69117 Heidelberg, Germany

[4] Pazmany Peter Catholic Univ, Fac Informat Technol & Bion, H-1083 Budapest, Hungary

[5] Heidelberg Univ, Dept Infect Dis, Virol, D-69120 Heidelberg, Germany

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

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2016年 / 113卷 / 13期

关键词：

light-sheet microscopy; RESOLFT; optical nanoscopy; 3D; live-cell imaging; PLANE ILLUMINATION MICROSCOPY; STIMULATED-EMISSION; LIVING CELLS; RESOLUTION; PROTEIN; SPIM; SPECIMENS; NANOSCOPY; BESSEL; BEAMS;

D O I：

10.1073/pnas.1522292113

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 present a plane-scanning RESOLFT [reversible saturable/switchable optical (fluorescence) transitions] light-sheet (LS) nanoscope, which fundamentally overcomes the diffraction barrier in the axial direction via confinement of the fluorescent molecular state to a sheet of subdiffraction thickness around the focal plane. To this end, reversibly switchable fluorophores located right above and below the focal plane are transferred to a nonfluorescent state at each scanning step. LS-RESOLFT nanoscopy offers wide-field 3D imaging of living biological specimens with low light dose and axial resolution far beyond the diffraction barrier. We demonstrate optical sections that are thinner by 5-12-fold compared with their conventional diffraction-limited LS analogs.

引用

页码：3442 / 3446

页数：5

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