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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 21, Iss. 13 — Jul. 1, 2013
  • pp: 15912–15925

Resolution and contrast enhancement in laser scanning microscopy using dark beam imaging

Harold Dehez, Michel Piché, and Yves De Koninck  »View Author Affiliations

Optics Express, Vol. 21, Issue 13, pp. 15912-15925 (2013)

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Laser scanning microscopy allows for three-dimensional imaging of cells with molecular specific labeling. However the spatial resolution of optical microscopy is fundamentally limited by the diffraction of light. In the last two decades many techniques have been introduced to enhance the resolution of laser scanning microscopes. However most of these techniques impose strong constraints on the specimen or rely on complex optical systems. These constraints limit the applicability of resolution improvement to various imaging modalities and sample types. To overcome these limitations, we introduce here a novel approach, which we called Switching LAser Mode (SLAM) microscopy, to enhance resolution and contrast in laser scanning microscopy. SLAM microscopy relies on subtracting images obtained with dark and bright modes, and exploits the smaller dimensions of the dark spot of the azimuthally polarized TE01 mode. With this approach, resolution is improved by a factor of two in confocal microscopy. The technique is not based on complex nonlinear processes and thus requires laser power similar to that used in conventional imaging, minimizing photo-damage. The flexibility of the approach enables retrofitting in commercial confocal and two-photon microscopes and opens avenues for resolution enhancement in fluorescence-independent microscopy.

© 2013 OSA

OCIS Codes
(140.3300) Lasers and laser optics : Laser beam shaping
(180.1790) Microscopy : Confocal microscopy
(180.5810) Microscopy : Scanning microscopy
(350.5730) Other areas of optics : Resolution
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: April 1, 2013
Revised Manuscript: May 31, 2013
Manuscript Accepted: May 31, 2013
Published: June 25, 2013

Virtual Issues
Vol. 8, Iss. 8 Virtual Journal for Biomedical Optics

Harold Dehez, Michel Piché, and Yves De Koninck, "Resolution and contrast enhancement in laser scanning microscopy using dark beam imaging," Opt. Express 21, 15912-15925 (2013)

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