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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 21, Iss. 22 — Nov. 4, 2013
  • pp: 26162–26173

Full-field dual-color 100-nm super-resolution imaging reveals organization and dynamics of mitochondrial and ER networks

Maia Brunstein, Kai Wicker, Karine Hérault, Rainer Heintzmann, and Martin Oheim  »View Author Affiliations

Optics Express, Vol. 21, Issue 22, pp. 26162-26173 (2013)

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Most structured illumination microscopes use a physical or syn–thetic grating that is projected into the sample plane to generate a periodic illumination pattern. Albeit simple and cost-effective, this arrangement hampers fast or multi-color acquisition, which is a critical requirement for time-lapse imaging of cellular and sub-cellular dynamics. In this study, we designed and implemented an interferometric approach allowing large-field, fast, dual-color imaging at an isotropic 100-nm resolution based on a sub-diffraction fringe pattern generated by the interference of two colliding evanescent waves. Our all-mirror-based system generates illumination pat-terns of arbitrary orientation and period, limited only by the illumination aperture (NA = 1.45), the response time of a fast, piezo-driven tip-tilt mirror (10 ms) and the available fluorescence signal. At low µW laser powers suitable for long-period observation of life cells and with a camera exposure time of 20 ms, our system permits the acquisition of super-resolved 50 µm by 50 µm images at 3.3 Hz. The possibility it offers for rapidly adjusting the pattern between images is particularly advantageous for experiments that require multi-scale and multi-color information. We demonstrate the performance of our instrument by imaging mitochondrial dynamics in cultured cortical astrocytes. As an illustration of dual-color excitation dual-color detection, we also resolve interaction sites between near-membrane mitochondria and the endoplasmic reticulum. Our TIRF-SIM microscope provides a versatile, compact and cost-effective arrangement for super-resolution imaging, allowing the investigation of co-localization and dynamic interactions between organelles – important questions in both cell biology and neurophysiology

© 2013 Optical Society of America

OCIS Codes
(100.6640) Image processing : Superresolution
(180.0180) Microscopy : Microscopy
(180.2520) Microscopy : Fluorescence microscopy
(180.3170) Microscopy : Interference microscopy

ToC Category:

Original Manuscript: August 27, 2013
Revised Manuscript: October 7, 2013
Manuscript Accepted: October 8, 2013
Published: October 24, 2013

Virtual Issues
Vol. 9, Iss. 1 Virtual Journal for Biomedical Optics

Maia Brunstein, Kai Wicker, Karine Hérault, Rainer Heintzmann, and Martin Oheim, "Full-field dual-color 100-nm super-resolution imaging reveals organization and dynamics of mitochondrial and ER networks," Opt. Express 21, 26162-26173 (2013)

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