By precisely managing fiber-optic nonlinearity with anomalous dispersion, we have demonstrated the control of generating plural few-optical-cycle pulses based on a 24-MHz Chromium:forsterite laser, allowing multicolor two-photon tissue imaging by wavelength mixing. The formation of high-order soliton and its efficient coupling to dispersive wave generation leads to phase-matched spectral broadening, and we have obtained a broadband continuum ranging from 830 nm to 1200 nm, delivering 5-nJ pulses with a pulse width of 10.5 fs using a piece of large-mode-area fiber. We locate the spectral enhancement at around 920 nm for the two-photon excitation of green fluorophores, and we can easily compress the resulting pulse close to its limited duration without the need for active pulse shaping. To optimize the wavelength mixing for sum-frequency excitation, we have realized the management of the power ratio and group delay between the soliton and dispersive wave by varying the initial pulse energy without additional delay control. We have thus demonstrated simultaneous three-color two-photon tissue imaging with contrast management between different signals. Our source optimization leads to efficient two-photon excitation reaching a 500-µm imaging depth under a low 14-mW illumination power. We believe our source development leads to an efficient and compact approach for driving multicolor two-photon fluorescence microscopy and other ultrafast investigations, such as strong-field-driven applications.
Generating ultrafast pulses with better spectrotemporal control is crucial for optimizing and characterizing nonlinear light–matter responses, yet it is limited by the gain bandwidth of laser media or the phase‐matching geometry of nonlinear processes. This work proposes a simple approach to independently manage a femtosecond source's spectral location and bandwidth. Self‐phase‐modulation‐enabled spectral broadening is first analyzed, which is potentially energy‐scalable using hollow‐core capillaries or multipass cells. It is demonstrated that the outmost lobes in the broadened spectrum show different dependencies on the initial pulse energy and duration. A simple yet effective toy model is introduced that successfully predicts broadband spectral tuning, and the impact of other nonlinear effects, dispersion, and input pulse asymmetry on the experimental scenario is also discussed. Thus a fiber‐based versatile source is demonstrated, which is compressible down to its transform‐limit duration, as short as 12.2 fs centered at 920 nm. In addition, bandwidth‐dependent third‐harmonic generation spectroscopy is performed from a dielectric metasurface with an optimized nonlinear response, and the dependency of laser bandwidth and pulse duration is investigated on the signal‐to‐background ratio of two‐photon images. It is believed that this demonstration will advance the investigation of bandwidth‐dependent nonlinear spectroscopy and microscopy.
We have demonstrated 30-Hz three-photon imaging using a single 24-MHz mode-locked Cr:forsterite oscillator with a center wavelength at 1260 nm. By managing the dispersion distribution in the resonator using double-chirped mirrors, we have produced 32-fs pulses with 22-nJ pulse energy. Using the oscillator as a driving source, we have realized multi-color three-photon images using a GFP-labeled Drosophila brain and an AF647-labeled mouse brain. To demonstrate the capability of deep-tissue imaging, we have obtained a 10-times higher SBR from the three-photon images than the two-photon results at different depths in a GFP-labeled Drosophila brain dissection. Furthermore, we have shown the impact of excitation pulse width on three-photon deep-tissue imaging. Our results indicate the superiority of using shorter pulses for deeper-tissue imaging, especially in the Drosophila brain. In addition, we have recorded the three-photon calcium imaging in vivo from the Drosophila mushroom body in response to external electric shocks. We believe our demonstration provides a robust approach for high-speed three-photon microscopy applications, especially for intravital investigations in the Drosophila brain.
Using a 24-MHz Cr:forsterite oscillator and the precisely controlled fiber-optic nonlinearity, we have simultaneously demonstrated sub-megawatt-peak-power femtosecond pulses at 1.3µm and compressible blue-shifted octave-spanning spectra.
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