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Proceedings Paper

Laser scanning stereomicroscopy for fast volumetric imaging with two-photon excitation and scanned Bessel beams
Author(s): Yanlong Yang; Xing Zhou; Runze Li; Mark Van Horn; Tong Peng; Ming Lei; Di Wu; Xun Chen; Baoli Yao; Tong Ye
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Paper Abstract

Bessel beams have been used in many applications due to their unique optical properties of maintaining their intensity profiles unchanged during propagation. In imaging applications, Bessel beams have been successfully used to provide extended focuses for volumetric imaging and uniformed illumination plane in light-sheet microscopy. Coupled with two-photon excitation, Bessel beams have been successfully used in realizing fluorescence projected volumetric imaging. We demonstrated previously a stereoscopic solution–two-photon fluorescence stereomicroscopy (TPFSM)–for recovering the depth information in volumetric imaging with Bessel beams. In TPFSM, tilted Bessel beams were used to generate stereoscopic images on a laser scanning two-photon fluorescence microscope; upon post image processing we could successfully provide 3D perception of acquired volume images by wearing anaglyph 3D glasses. However, tilted Bessel beams were generated by shifting either an axicon or an objective laterally; the slow imaging speed and severe aberrations made it hard to use in real-time volume imaging. In this article, we report recent improvements of TPFSM with newly designed scanner and imaging software, which allows 3D stereoscopic imaging without moving any of the optical components on the setup. This improvement has dramatically improved focusing qualities and imaging speed so that the TPFSM can be performed potentially in real-time to provide 3D visualization in scattering media without post image processing.

Paper Details

Date Published: 5 March 2015
PDF: 4 pages
Proc. SPIE 9329, Multiphoton Microscopy in the Biomedical Sciences XV, 93292W (5 March 2015); doi: 10.1117/12.2079646
Show Author Affiliations
Yanlong Yang, Xi'an Institute of Optics and Precision Mechanics (China)
Clemson Univ. (United States)
Xing Zhou, Xi'an Institute of Optics and Precision Mechanics (China)
Runze Li, Xi'an Institute of Optics and Precision Mechanics (China)
Mark Van Horn, Medical Univ. of South Carolina (United States)
Tong Peng, Xi’an Institute of Optics and Precision Mechanics (China)
Ming Lei, Xi’an Institute of Optics and Precision Mechanics (China)
Di Wu, Xi’an Institute of Optics and Precision Mechanics (China)
Xun Chen, Clemson Univ. (United States)
Baoli Yao, Xi’an Institute of Optics and Precision Mechanics (China)
Tong Ye, Clemson Univ. (United States)


Published in SPIE Proceedings Vol. 9329:
Multiphoton Microscopy in the Biomedical Sciences XV
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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