摘要： Multiphoton microscopy (MPM) is an important bio-imaging tool. Different modalities can serve as a contrast agent, such as second-/third-harmonic generation (SHG/THG) and two-/three-photon excitation fluorescence (2PEF/3PEF). Ultrafast lasers with flexible wavelength tunability are crucial for driving MPM bio-imaging, and the conventional solution relies on ultrafast Ti:sapphire lasers plus an optical parametric oscillator/amplifier. Recently, we have demonstrated that ultrafast fiber lasers are a potential solution to implementing compact, robust, and wavelength tunable femtosecond sources for driving MPM. To realize wavelength tunability we employ self-phase modulation (SPM) in optical fibers to broaden a narrowband input spectrum of Yb-/Er-doped fiber lasers (YDFLs/EDFLs) up to >400-nm wide with well-isolated spectral lobes; filtering the leftmost/rightmost lobes leads to nearly transform-limited pulses [1–6]. Such a SPM-enabled spectral selection (SESS) allows us to obtain wavelength widely tunable femtosecond pulses for MPM [2,5,6]. In this submission, we representatively demonstrate label-free harmonic generation microscopy (HGM) in human skin and brain tissues. Figure 1(a) depicts a scanning microscope driven by an EDFL-based SESS source. The EDFL operates at 31-MHz repetition rate and generates 290-fs pulses centered at 1550 nm. The narrowband EDFL [blue curve in Fig. 1(a)] is coupled into 9-cm optical fiber (10-μm mode-field diameter and -10 fs2/mm group-velocity dispersion at 1550 nm). The output spectrum is shown as the red curve in Fig. 1(b) under 85-nJ coupled pulse energy. We use optical filters to select the leftmost spectral lobe peaking at 1250 nm, which leads to 11.7-nJ, 47-fs pulses. Then we employ these pulses to drive a scanning microscope and conduct HGM in human skin and brain tissues. Figure 1(c) shows the dermal papilla at the junction of epidermis and upper dermis in human skin. Basal cells are visualized by THG (cyan hot) due to optical inhomogeneity at the interface (e.g., cell membrane); SHG (red hot) originates from the non-centrosymmetric structure of collagen fibers. In Fig. 1(d), neural network and brain vasculature in human brain tissue can be visualized by THG and SHG, respectively [Fig. 1(d)]. THG contrast inside the vasculature shows also red blood cells. In conclusion, we report on MPM deep-tissue imaging enabled by an EDFL-based SESS source. It is noteworthy that besides HGM excited by 1250-nm femtosecond pulses demonstrated here, the SESS source also supports 1300-/1700-nm illumination for 3PEF of green/red fluorescent protein (GFP/RFP) [7,8]. Such a fiber-based solution can be applied to many important applications, such as histopathology, morphology, and neuroscience.