help button home button Biophys. J.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Biophys. J. BioFAST: First Published March 7, 2008. doi:10.1529/biophysj.107.120345
© 2008 by the Biophysical Society.

A more recent version of this article appeared on June 15, 2008.
This Article
Right arrow Full Text (Rapid PDF)
Right arrow Supplement
Right arrow All Versions of this Article:
biophysj.107.120345v1
biophysj.107.120345v2
94/12/4957    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gustafsson, M. G.L.
Right arrow Articles by Sedat, J. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gustafsson, M. G.L.
Right arrow Articles by Sedat, J. W.

SPECTROSCOPY, IMAGING, OTHER TECHNIQUES

Three-dimensional Resolution Doubling in Widefield Fluorescence Microscopy by Structured Illumination

Mats G.L. Gustafsson 1*, Lin Shao 1, Peter M. Carlton 1, C. J. Rachel Wang 2, Inna N. Golubovskaya 2, W. Zacheus Cande 2, David A. Agard 1 and John W. Sedat 1

1 University of California, San Francisco
2 University of California, Berkeley

* To whom correspondence should be addressed. E-mail: mats{at}msg.ucsf.edu.

Submitted on August 24, 2007
Revised on September 26, 2007
Accepted on 6 November 2007


  Abstract
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 widefield 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.

Key Words: HELM, cytoskeleton, microtubules, patterned excitation, super-resolution, synaptonemal complex




This article has been cited by other articles:


Home page
 ScienceHome page
L. Schermelleh, P. M. Carlton, S. Haase, L. Shao, L. Winoto, P. Kner, B. Burke, M. C. Cardoso, D. A. Agard, M. G. L. Gustafsson, et al.
Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy
Science, June 6, 2008; 320(5881): 1332 - 1336.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Copyright © 2008 by the Biophysical Society.