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Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy

Yu, Jiun-Yann and Kuo, Chun-Hung and Holland, Daniel B. and Chen, Yenyu and Ouyang, Mingxing and Blake, Geoffrey A. and Zadoyan, Ruben and Guo, Chin-Lin (2011) Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy. Journal of Biomedical Optics, 16 (11). Art. No. 116009. ISSN 1083-3668.

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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.

Item Type:Article
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Blake, Geoffrey A.0000-0003-0787-1610
Additional Information:© 2011 Society of Photo-Optical Instrumentation Engineers (SPIE). Paper 11137RR received Mar. 22, 2011; revised manuscript received Sep. 3, 2011; accepted for publication Sep. 19, 2011; published online Nov. 3, 2011. This work is a collaborative research effort of the Cell Polarity Laboratory at the California Institute of Technology and the Technology and Applications Center at Newport Corporation. The authors would like to thank Mr. Craig Goldberg of Newport Corporation for facilitating the collaboration, and Professor Shi-Wei Chu for the inspiring discussions. Mr. Ji Hun Kim of Caltech is sincerely acknowledged for the measurement of the surface profile of the optical diffuser. CLG recognizes support from the Ellison Medical Foundation and the Weston Havens Foundation. Support from the National Science Foundation Chemistry Research Instrumentation and Facilities: Instrument Development program to GAB is gratefully acknowledged.
Funding AgencyGrant Number
Ellison Medical FoundationUNSPECIFIED
Weston Havens FoundationUNSPECIFIED
Subject Keywords:microscopy; multiphoton processes; diffusers; imaging systems. amplifiers, biological tissues, biomedical optical imaging, cellular biophysics, fluorescence, high-speed optical techniques, image resolution, laser applications in medicine, molecular biophysics, multiphoton processes, optical focusing, optical microscopy, proteins
Issue or Number:11
Classification Code:PACS: 87.63.It;; 87.17.-d; 87.14.E-;;
Record Number:CaltechAUTHORS:20120106-140226269
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Official Citation:Jiun-Yann Yu, Chun-Hung Kuo, Daniel B. Holland, Yenyu Chen, Mingxing Ouyang, Geoffrey A. Blake, Ruben Zadoyan and Chin-Lin Guo, "Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy", J. Biomed. Opt. 16, 116009 (Nov 03, 2011); doi:10.1117/1.3647570
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:28705
Deposited By: Ruth Sustaita
Deposited On:06 Jan 2012 22:37
Last Modified:03 Oct 2019 03:34

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