Simple approach to three-color two-photon microscopy by a fiber-optic wavelength convertor

Kuen Che Li, Lynn L.H. Huang, Jhih Hao Liang, Ming-Che Chan*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

11 Scopus citations


A simple approach to multi-color two-photon microscopy of the red, green, and blue fluorescent indicators was reported based on an ultra-compact 1.03-μm femtosecond laser and a nonlinear fiber. Inside the nonlinear fiber, the 1.03-μm laser pulses were simultaneously blue-shifted to 0.6~0.8 μm and red-shifted to 1.2~1.4 μm region by the Cherenkov radiation and fiber Raman gain effects. The wavelength-shifted 0.6~0.8 μm and 1.2~1.4 μm radiations were co-propagated with the residual non-converted 1.03-μm pulses inside the same nonlinear fiber to form a fiber-output three-color femtosecond source. The application of the multi-wavelength sources on multi-color two-photon fluorescence microscopy were also demonstrated. Overall, due to simple system configuration, convenient wavelength conversion, easy wavelength tunability within the entire 0.7~1.35 μm biopenetration window and less requirement for high power and bulky light sources, the simple approach to multi-color two-photon microscopy could be widely applicable as an easily implemented and excellent research tool for future biomedical and possibly even clinical applications.

Original languageEnglish
Article number#274506
Pages (from-to)4803-4815
Number of pages13
JournalBiomedical Optics Express
Issue number11
StatePublished - 1 Nov 2016


  • Fluorescence microscopy
  • Multispectral and hyperspectral imaging
  • Nonlinear microscopy
  • Nonlinear optics, fibers
  • Ultrafast nonlinear optics

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