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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 21, Iss. 5 — Mar. 11, 2013
  • pp: 5998–6008

Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application

Zeno Lavagnino, Francesca Cella Zanacchi, Emiliano Ronzitti, and Alberto Diaspro  »View Author Affiliations


Optics Express, Vol. 21, Issue 5, pp. 5998-6008 (2013)
http://dx.doi.org/10.1364/OE.21.005998


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Abstract

In this work we report the advantages provided by two photon excitation (2PE) implemented in a selective plane illumination microscopy (SPIM) when imaging thick scattering samples. In particular, a detailed analysis of the effects induced on the real light sheet excitation intensity distribution is performed. The comparison between single-photon and two-photon excitation profiles shows the reduction of the scattering effects and sample-induced aberrations provided by 2PE-SPIM. Furthermore, uniformity of the excitation distribution and the consequent improved image contrast is shown when imaging scattering phantom samples in depth by 2PE-SPIM. These results show the advantages of 2PE-SPIM and suggest how this combination can further enhance the SPIM performance. Phantom samples have been designed with optical properties compatible with biological applications of interest.

© 2013 OSA

OCIS Codes
(180.0180) Microscopy : Microscopy
(110.0113) Imaging systems : Imaging through turbid media

ToC Category:
Microscopy

History
Original Manuscript: November 29, 2012
Revised Manuscript: January 26, 2013
Manuscript Accepted: February 16, 2013
Published: March 4, 2013

Virtual Issues
Vol. 8, Iss. 4 Virtual Journal for Biomedical Optics

Citation
Zeno Lavagnino, Francesca Cella Zanacchi, Emiliano Ronzitti, and Alberto Diaspro, "Two-photon excitation selective plane illumination microscopy (2PE-SPIM) of highly scattering samples: characterization and application," Opt. Express 21, 5998-6008 (2013)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-21-5-5998


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