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Applied Optics

Applied Optics


  • Editor: Joseph N. Mait
  • Vol. 49, Iss. 32 — Nov. 10, 2010
  • pp: 6348–6353

Two-photon fluorescence excitation spectroscopy by pulse shaping ultrabroad-bandwidth femtosecond laser pulses

Bingwei Xu, Yves Coello, Vadim V. Lozovoy, and Marcos Dantus  »View Author Affiliations

Applied Optics, Vol. 49, Issue 32, pp. 6348-6353 (2010)

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A fast and automated approach to measuring two-photon fluorescence excitation (TPE) spectra of fluorophores with high resolution ( 2 nm ) by pulse shaping ultrabroad-bandwidth femtosecond laser pulses is demonstrated. Selective excitation in the range of 675 990 nm was achieved by imposing a series of specially designed phase and amplitude masks on the excitation pulses using a pulse shaper. The method eliminates the need for laser tuning and is, thus, suitable for non-laser-expert use. The TPE spectrum of Fluorescein was compared with independent measurements and the spectra of the pH-sensitive dye 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) in acidic and basic environments were measured for the first time using this approach.

© 2010 Optical Society of America

OCIS Codes
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(190.7110) Nonlinear optics : Ultrafast nonlinear optics
(300.6410) Spectroscopy : Spectroscopy, multiphoton
(320.5540) Ultrafast optics : Pulse shaping

ToC Category:

Original Manuscript: July 20, 2010
Revised Manuscript: October 10, 2010
Manuscript Accepted: October 12, 2010
Published: November 9, 2010

Virtual Issues
Vol. 6, Iss. 1 Virtual Journal for Biomedical Optics

Bingwei Xu, Yves Coello, Vadim V. Lozovoy, and Marcos Dantus, "Two-photon fluorescence excitation spectroscopy by pulse shaping ultrabroad-bandwidth femtosecond laser pulses," Appl. Opt. 49, 6348-6353 (2010)

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