Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination

被引：1178

作者：

Gustafsson, Mats G. L. ^{[1
,2
]}

Shao, Lin ^{[3
,4
]}

Carlton, Peter M. ^{[3
,4
]}

Wang, C. J. Rachel ^{[5
]}

Golubovskaya, Inna N. ^{[5
]}

Cande, W. Zacheus ^{[5
]}

Agard, David A. ^{[3
,4
,6
]}

Sedat, John W. ^{[3
,4
]}

机构：

[1] Univ Calif San Francisco, Dept Physiol, San Francisco, CA 94143 USA

[2] Univ Calif San Francisco, Bioengn Program, San Francisco, CA 94143 USA

[3] Univ Calif San Francisco, Keck Adv Microscopy Lab, San Francisco, CA 94143 USA

[4] Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94143 USA

[5] Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA

[6] Howard Hughes Med Inst, Chevy Chase, MD 20815 USA

来源：

BIOPHYSICAL JOURNAL | 2008年 / 94卷 / 12期

关键词：

D O I：

10.1529/biophysj.107.120345

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

Structured illumination microscopy is a method that can increase the spatial resolution of wide-field fluorescence microscopy beyond its classical limit by using spatially structured illumination light. Here we describe how this method can be applied in three dimensions to double the axial as well as the lateral resolution, with true optical sectioning. A grating is used to generate three mutually coherent light beams, which interfere in the specimen to form an illumination pattern that varies both laterally and axially. The spatially structured excitation intensity causes normally unreachable high-resolution information to become encoded into the observed images through spatial frequency mixing. This new information is computationally extracted and used to generate a three-dimensional reconstruction with twice as high resolution, in all three dimensions, as is possible in a conventional wide-field microscope. The method has been demonstrated on both test objects and biological specimens, and has produced the first light microscopy images of the synaptonemal complex in which the lateral elements are clearly resolved.

引用

页码：4957 / 4970

页数：14

共 33 条

[21] Laterally modulated excitation microscopy: Improvement of resolution by using a diffraction grating
Heintzmann, R
Cremer, C
[J]. OPTICAL BIOPSIES AND MICROSCOPIC TECHNIQUES III, PROCEEDINGS OF, 1999, 3568 : 185 - 196
[22] Saturated patterned excitation microscopy - a concept for optical resolution improvement
Heintzmann, R
Jovin, TM
Cremer, C
[J]. JOURNAL OF THE OPTICAL SOCIETY OF AMERICA A-OPTICS IMAGE SCIENCE AND VISION, 2002, 19 (08): : 1599 - 1609
[23] Ultra-high resolution imaging by fluorescence photoactivation localization microscopy
Hess, Samuel T.
Girirajan, Thanu P. K.
Mason, Michael D.
[J]. BIOPHYSICAL JOURNAL, 2006, 91 (11) : 4258 - 4272
[24] Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission
Klar, TA
Jakobs, S
Dyba, M
Egner, A
Hell, SW
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (15) : 8206 - 8210
[25] Real time 3D fluorescence microscopy by two beam interference illumination
Neil, MAA
Juskaitis, R
Wilson, T
[J]. OPTICS COMMUNICATIONS, 1998, 153 (1-3) : 1 - 4
[26] Method of obtaining optical sectioning by using structured light in a conventional microscope
Neil, MAA
Juskaitis, R
Wilson, T
[J]. OPTICS LETTERS, 1997, 22 (24) : 1905 - 1907
[27] The genetics and molecular biology of the synaptonemal complex
Page, SL
Hawley, RS
[J]. ANNUAL REVIEW OF CELL AND DEVELOPMENTAL BIOLOGY, 2004, 20 : 525 - 558
[28] Pawley J.B., 2006, HDB BIOL CONFOCAL MI
[29] Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM)
Rust, Michael J.
Bates, Mark
Zhuang, Xiaowei
[J]. NATURE METHODS, 2006, 3 (10) : 793 - 795
[30] Wide-field subdiffraction RESOLFT microscopy using fluorescent protein photoswitching
Schwentker, Miriam A.
Bock, Hannes
Hofmann, Michael
Jakobs, Stefan
Bewersdorf, Joerg
Eggeling, Christian
Hell, Stefan W.
[J]. MICROSCOPY RESEARCH AND TECHNIQUE, 2007, 70 (03) : 269 - 280

← 1 2 3 4 →