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, Leilei Peng

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1296-1308
Number of pages13
JournalBiomedical Optics Express
Volume5
Issue number5
DOIs
StatePublished - May 1 2014

ASJC Scopus subject areas

  • Biotechnology
  • Atomic and Molecular Physics, and Optics

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