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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 5, Iss. 5 — May. 1, 2014
  • pp: 1296–1308

Cellular imaging of deep organ using two-photon Bessel light-sheet nonlinear structured illumination microscopy

Ming Zhao, Han Zhang, Yu Li, Amit Ashok, Rongguang Liang, Weibin Zhou, and Leilei Peng  »View Author Affiliations

Biomedical Optics Express, Vol. 5, Issue 5, pp. 1296-1308 (2014)

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In vivo fluorescent cellular imaging of deep internal organs is highly challenging, because the excitation needs to penetrate through strong scattering tissue and the emission signal is degraded significantly by photon diffusion induced by tissue-scattering. We report that by combining two-photon Bessel light-sheet microscopy with nonlinear structured illumination microscopy (SIM), live samples up to 600 microns wide can be imaged by light-sheet microscopy with 500 microns penetration depth, and diffused background in deep tissue light-sheet imaging can be reduced to obtain clear images at cellular resolution in depth beyond 200 microns. We demonstrate in vivo two-color imaging of pronephric glomeruli and vasculature of zebrafish kidney, whose cellular structures located at the center of the fish body are revealed in high clarity by two-color two-photon Bessel light-sheet SIM.

© 2014 Optical Society of America

OCIS Codes
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.6900) Medical optics and biotechnology : Three-dimensional microscopy

ToC Category:

Original Manuscript: February 21, 2014
Revised Manuscript: March 24, 2014
Manuscript Accepted: March 24, 2014
Published: March 31, 2014

Ming Zhao, Han Zhang, Yu Li, Amit Ashok, Rongguang Liang, Weibin Zhou, and Leilei Peng, "Cellular imaging of deep organ using two-photon Bessel light-sheet nonlinear structured illumination microscopy," Biomed. Opt. Express 5, 1296-1308 (2014)

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