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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 9 — Sep. 1, 2012
  • pp: 1972–1977

Three-color femtosecond source for simultaneous excitation of three fluorescent proteins in two-photon fluorescence microscopy

Ke Wang, Tzu-Ming Liu, Juwell Wu, Nicholas G. Horton, Charles P. Lin, and Chris Xu  »View Author Affiliations

Biomedical Optics Express, Vol. 3, Issue 9, pp. 1972-1977 (2012)

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We demonstrate a fiber-based, three-color femtosecond source for simultaneous imaging of three fluorescent proteins (FPs) using two-photon fluorescence microscopy (2PM). The three excitation wavelengths at 775 nm, 864 nm and 950 nm, are obtained through second harmonic generation (SHG) of the 1550-nm pump laser and the 1728-nm and 1900-nm solitons generated through soliton self-frequency shift (SSFS) in a large-mode-area (LMA) fiber. These energetic pulses are well matched to the two-photon excitation peaks of red, cyan and yellow fluorescent proteins (TagRFPs, TagCFPs, and TagYFPs) for efficient excitation. We demonstrate simultaneous 2PM of human melanoma cells expressing a “rainbow” combination of these three fluorescent proteins.

© 2012 OSA

OCIS Codes
(060.4370) Fiber optics and optical communications : Nonlinear optics, fibers
(180.2520) Microscopy : Fluorescence microscopy
(190.2620) Nonlinear optics : Harmonic generation and mixing
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: June 28, 2012
Revised Manuscript: July 25, 2012
Manuscript Accepted: July 25, 2012
Published: July 31, 2012

Ke Wang, Tzu-Ming Liu, Juwell Wu, Nicholas G. Horton, Charles P. Lin, and Chris Xu, "Three-color femtosecond source for simultaneous excitation of three fluorescent proteins in two-photon fluorescence microscopy," Biomed. Opt. Express 3, 1972-1977 (2012)

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