Real-time monitoring the variation of chlorophyll distributions and cellular structures in leaves during plant growth provides important information for understanding the physiological statuses of plants. Two-photonexcited autofluorescence imaging and second harmonic generation imaging of leaves can be used for monitoring the nature intrinsic fluorophores distribution and cellular structures of leaves by the use of the near-infrared region of light which has minimal light absorption by endogenous molecules and thus increases tissue penetration. However, the two-photon absorption peak of intrinsic fluorophores of a ficus benjamina leaf is 50 nm away from the second harmonic generation excitation wavelength, which cannot be effectively excited by a femtosecond laser beam with one central wavelength. This paper shows that a highly polarized supercontinuum light generated from a birefringent nonlinear photonic crystal fiber with two zero-dispersion wavelengths can effectively excite two-photon autofluorescence as well as second harmonic generation signals for simultaneously monitoring intrinsic fluorophore distributions and non-centrosymmetric structures of leaves.
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