Multi-Spectral Two-Photon Excited Fluorescence Microscopy Using Supercontinuum Light Source

We report on a novel technique of multi-spectral two-photon-excited fluorescence microscopy by use of broadband supercontinuum light. A supercontinuum in the near-infrared region is generated in a 4.5-mm-long photonic crystal fiber using a Ti:sapphire femtosecond oscillator. The supercontinuum is used as a multi-spectral excitation light source in a two-photon excited fluorescence microscope. We demonstrate that three-color fluorescence images of organelles in a cell can be simultaneously acquired.

[1]  W. Webb,et al.  Multiphoton fluorescence excitation: new spectral windows for biological nonlinear microscopy. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Govind P. Agrawal,et al.  Nonlinear Fiber Optics , 1989 .

[3]  J. Thøgersen,et al.  Coherent anti-Stokes Raman scattering microscopy with a photonic crystal fiber based light source , 2003, Conference on Lasers and Electro-Optics, 2003. CLEO '03..