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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 9, Iss. 3 — Mar. 6, 2014

An adaptive approach for uniform scanning in multifocal multiphoton microscopy with a spatial light modulator

Naoya Matsumoto, Shigetoshi Okazaki, Yasuko Fukushi, Hisayoshi Takamoto, Takashi Inoue, and Susumu Terakawa  »View Author Affiliations

Optics Express, Vol. 22, Issue 1, pp. 633-645 (2014)

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We propose high-quality generation of uniform multiple fluorescence spots (MFS) with a spatial light modulator (SLM) and demonstrate uniform laser scanning in multifocal multiphoton microscopy (MMM). The MFS excitation method iteratively updates a computer-generated hologram (CGH) using correction coefficients to improve the fluorescence intensity distribution in a dye solution whose consistency is uniform. This simple correction method can be applied for calibration of the MMM before observation of living tissue. We experimentally demonstrate an improvement of the uniformity of a 10 × 10 grid of MFS by using a dye solution. After the calibration, we performed laser scanning with two-photon excitation to observe fluorescent polystyrene beads, as well as the gastric gland of a guinea pig specimen.

© 2014 Optical Society of America

OCIS Codes
(090.1760) Holography : Computer holography
(180.2520) Microscopy : Fluorescence microscopy
(180.5810) Microscopy : Scanning microscopy
(230.3720) Optical devices : Liquid-crystal devices
(230.6120) Optical devices : Spatial light modulators
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: August 2, 2013
Revised Manuscript: December 11, 2013
Manuscript Accepted: December 17, 2013
Published: January 6, 2014

Virtual Issues
Vol. 9, Iss. 3 Virtual Journal for Biomedical Optics

Naoya Matsumoto, Shigetoshi Okazaki, Yasuko Fukushi, Hisayoshi Takamoto, Takashi Inoue, and Susumu Terakawa, "An adaptive approach for uniform scanning in multifocal multiphoton microscopy with a spatial light modulator," Opt. Express 22, 633-645 (2014)

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