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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 17, Iss. 16 — Aug. 3, 2009
  • pp: 13737–13746

Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses

Keisuke Isobe, Akira Suda, Masahiro Tanaka, Fumihiko Kannari, Hiroyuki Kawano, Hideaki Mizuno, Atsushi Miyawaki, and Katsumi Midorikawa  »View Author Affiliations

Optics Express, Vol. 17, Issue 16, pp. 13737-13746 (2009)

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We propose two-photon excited fluorescence (TPEF) microscopy employing a novel phase modulation technique of ultra-broadband laser pulses, which allows the relative excitation of an individual fluorophore with respect to other fluorophores. This technique is based on the generation of multi-wavelength pulse train, which independently interacts with each fluorophore. Our technique is applied to dual-color imaging of cells expressing two types of fluorescent proteins. We achieve the selective excitation of one over the other and vice versa. The product of the maximum contrast ratios exceeds 100. We also demonstrate yielded equal excitation rates in the simultaneous excitation. By the selective excitation of a donor fluorescent protein, fluorescence resonance energy transfer imaging is also achieved.

© 2009 OSA

OCIS Codes
(120.5060) Instrumentation, measurement, and metrology : Phase modulation
(180.2520) Microscopy : Fluorescence microscopy
(190.4180) Nonlinear optics : Multiphoton processes
(320.5540) Ultrafast optics : Pulse shaping
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: May 21, 2009
Revised Manuscript: June 28, 2009
Manuscript Accepted: June 30, 2009
Published: July 24, 2009

Virtual Issues
Vol. 4, Iss. 10 Virtual Journal for Biomedical Optics

Keisuke Isobe, Akira Suda, Masahiro Tanaka, Fumihiko Kannari, Hiroyuki Kawano, Hideaki Mizuno, Atsushi Miyawaki, and Katsumi Midorikawa, "Multifarious control of two-photon excitation of multiple fluorophores achieved by phase modulation of ultra-broadband laser pulses," Opt. Express 17, 13737-13746 (2009)

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