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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 22, Iss. 6 — Mar. 24, 2014
  • pp: 7028–7039

Simultaneous dual-color 3D STED microscopy

Christian Osseforth, Jeffrey R. Moffitt, Lothar Schermelleh, and Jens Michaelis  »View Author Affiliations

Optics Express, Vol. 22, Issue 6, pp. 7028-7039 (2014)

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We describe the design and implementation of a stimulated emission depletion (STED) microscope which allows simultaneous three-dimensional super-resolution imaging in two colors. A super-continuum laser source is used to provide all spectral bands necessary for excitation and efficient depletion to achieve a lateral and axial resolution of ~35 nm and ~90 nm respectively. We characterize the systems' performance by imaging colloidal particles and single fluorescent molecules. Its biological applicability is demonstrated by dual-color imaging of nuclear pore complexes and of DNA replication sites in mammalian cells.

© 2014 Optical Society of America

OCIS Codes
(000.1430) General : Biology and medicine
(100.6640) Image processing : Superresolution
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(350.5730) Other areas of optics : Resolution

ToC Category:

Original Manuscript: January 9, 2014
Revised Manuscript: March 12, 2014
Manuscript Accepted: March 12, 2014
Published: March 18, 2014

Virtual Issues
Vol. 9, Iss. 5 Virtual Journal for Biomedical Optics

Christian Osseforth, Jeffrey R. Moffitt, Lothar Schermelleh, and Jens Michaelis, "Simultaneous dual-color 3D STED microscopy," Opt. Express 22, 7028-7039 (2014)

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