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Journal of Biomedical Optics • Open Access

Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy
Author(s): Jiun-Yann Yu; Mingxing Ouyang; Chin-Lin Guo; Chun-Hung Kuo; Ruben Zadoyan; Daniel B. Holland; Geoffrey A. Blake; Yenyu Chen

Paper Abstract

Optical sectioning provides three-dimensional (3D) information in biological tissues. However, most imaging techniques implemented with optical sectioning are either slow or deleterious to live tissues. Here, we present a simple design for wide-field multiphoton microscopy, which provides optical sectioning at a reasonable frame rate and with a biocompatible laser dosage. The underlying mechanism of optical sectioning is diffuser-based temporal focusing. Axial resolution comparable to confocal microscopy is theoretically derived and experimentally demonstrated. To achieve a reasonable frame rate without increasing the laser power, a low-repetition-rate ultrafast laser amplifier was used in our setup. A frame rate comparable to that of epifluorescence microscopy was demonstrated in the 3D imaging of fluorescent protein expressed in live epithelial cell clusters. In this report, our design displays the potential to be widely used for video-rate live-tissue and embryo imaging with axial resolution comparable to laser scanning microscopy.

Paper Details

Date Published: 1 November 2011
PDF: 10 pages
J. Biomed. Opt. 16(11) 116009 doi: 10.1117/1.3647570
Published in: Journal of Biomedical Optics Volume 16, Issue 11
Show Author Affiliations
Jiun-Yann Yu, California Institute of Technology (United States)
Mingxing Ouyang, California Institute of Technology (United States)
Chin-Lin Guo, California Institute of Technology (United States)
Chun-Hung Kuo, Newport Corp. (United States)
Ruben Zadoyan, Newport Corp. (United States)
Daniel B. Holland, California Institute of Technology (United States)
Geoffrey A. Blake, California Institute of Technology (United States)
Yenyu Chen, Stanford Univ. School of Medicine (United States)

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