Figure 1 shows the schematic of our PCF-based multimodal multiphoton NLO microspectroscopy imaging system. This microspectroscopy imaging modality was modified and developed from a previously described system.15,16 An LD-pumped green laser (Verdi V5, Coherent) was used as the pumping source of a home-made Ti:sapphire femtosecond oscillator. In this system, the output wavelength was centered at 800 nm with pulse duration of 60 fs, repetition rate of 56.1 MHz, and average output power of 460 mW. Figure 2(a) shows the spectral profile of laser oscillator, and the bandwidth of beams generated from oscillator is about 4 nm (full width half maximum). Figure 2(b) shows the measured pulse width of laser oscillator by autocorrelator, and the laser pulse duration is 60 fs with assuming Gaussian intensity profile. The femtosecond pulses passed through an optical isolator (IO-5-NIR-LP, Thorlabs, New Jersey), and the output power was divided into two beams (3:1) using a beam splitter (BP133, Thorlabs). The reflected beam was then transmitted through a series of optical components for spatial filtering, and narrow band “pump pulses” in CARS process, the pulse of 0.8 nm with the center wavelength 800 nm, was generated by combining two filters (LL01-808-12.5 and custom made filter by Semrock). Figure 2(c) shows the spectrum of our narrowband pump beam. In contrast, the transmitted pulse was coupled into the PCF (FemtoWHITE 800, NKT Photonics, Denmark) through an objective lens in order to make ultrabroadband “Stokes pulses.” Figure 2(d) shows the measured spectrum of Stokes beam. The narrowband pump and broadband Stokes pulses were spatially and temporally overlapped using a long wavelength pass filter (BLP01-785R-25, Semrock, Illinois), and the two beams were passed through galvanometric mirrors. The collinear beams were tightly focused into the sample using an objective lens (LU Plan Fluor, 0.9 NA, , Nikon, Japan), and forward nonlinear signals were collected by a condenser lens (0.4 NA, , Olympus, Japan). The sample was mounted on an -axis piezo translation stage (SLC-1730, SmarAct, Germany), and its -axis was controlled by a piezoelectric objective lens positioning system (Mipos100, Piezosystemjena, Germany). After the nonlinear signals passed through a short pass filter (FF01-775/SP-25, Semrock), the CARS spectrum signal was detected using a monochromator (DongWoo i500) and then recorded using a charge-coupled device camera (iDus, Newton) having an effective spectral resolution of .