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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 19 — Sep. 10, 2012
  • pp: 21805–21814

Scanned light sheet microscopy with confocal slit detection

Eugen Baumgart and Ulrich Kubitscheck  »View Author Affiliations


Optics Express, Vol. 20, Issue 19, pp. 21805-21814 (2012)
http://dx.doi.org/10.1364/OE.20.021805


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Abstract

In light sheet fluorescence microscopy optical sectioning is achieved by illuminating the sample orthogonally to the detection pathway with a thin, focused sheet of light. However, light scattering within the sample often deteriorates the optical sectioning effect. Here, we demonstrate that contrast and degree of confocality can greatly be increased by combining scanned light sheet fluorescence excitation and confocal slit detection. A high frame rate was achieved by using the “rolling shutter” of a scientific CMOS camera as a slit detector. Synchronizing the “rolling shutter” with the scanned illumination beam results in confocal line detection. Acquiring image data with selective plane illumination minimizes photo-damage while simultaneously enhancing contrast, optical sectioning and signal-to-noise ratio. Thus the imaging principle presented here merges the benefits of scanned light sheet microscopy and line-scanning confocal imaging.

© 2012 OSA

OCIS Codes
(170.1790) Medical optics and biotechnology : Confocal microscopy
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.2520) Microscopy : Fluorescence microscopy
(180.5810) Microscopy : Scanning microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Microscopy

History
Original Manuscript: July 20, 2012
Revised Manuscript: August 13, 2012
Manuscript Accepted: September 5, 2012
Published: September 7, 2012

Virtual Issues
Vol. 7, Iss. 11 Virtual Journal for Biomedical Optics

Citation
Eugen Baumgart and Ulrich Kubitscheck, "Scanned light sheet microscopy with confocal slit detection," Opt. Express 20, 21805-21814 (2012)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-20-19-21805


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