Two-photon fluorescence microscope with a hollow-core photonic crystal fiber

Since the first demonstration in 1990, two-photon fluorescence microscopy (TPFM) has made a great impact on biomedical researches. With its high penetration ability, low out-of-focus photodamage, and intrinsic three-dimensional (3D) sectioning capability, TPFM has been widely applied to various medical diagnosis and genome researches. Recently, single-mode optical fibers were introduced into the TPFM systems for remote optical pulse delivery. Fiber-based TPFM has advantages including isolating the vibration from laser and electronic devices, flexible system design, and low cross-talks. It is also the first step toward an all-fiber based two-photon endoscope. However, due to serious temporal broadening when conventional Ti:sapphire based femtosecond pulses propagate through the fiber, the two-photon excitation efficiency of the fiber-optic TPFM is much lower than the conventional one. The temporal broadening effect mainly comes from group velocity dispersion (GVD) and self-phase modulation (SPM), which also leads to significant spectral broadening. To reduce the temporal broadening effect, here we present a hollow-core photonic-bandgap fiber based TPFM. By replacing the conventional single-mode fiber with the hollow core photonic bandgap fiber, the GVD and SPM effects can be greatly reduced for high intensity, ultra-short pulse delivery. Femtosecond Ti:sapphire pulses passing through the fiber with negligible GVD and SPM effects is demonstrated in this paper. Much improvement of two-photon fluorescence excitation efficiency is thus achieved with the hollow-core photonic-bandgap fiber based TPFM.