An adaptive excitation source for high-speed multiphoton microscopy.

Optical imaging is important for understanding brain function. However, established methods with high spatiotemporal resolution are limited by the potential for laser damage to living tissues. We describe an adaptive femtosecond excitation source that only illuminates the region of interest, which leads to a 30-fold reduction in the power requirement for two- or three-photon imaging of brain activity in awake mice for improved high-speed longitudinal neuroimaging.

Investigation of the long wavelength limit of soliton self-frequency shift in a silica fiber.

We explore the long wavelength limit of soliton self-frequency shift in silica-based fibers experimentally and using numerical simulation. We found that the longest wavelength soliton generated by soliton self-frequency shift is approximately 2500 nm because the soliton loses its energy rapidly at wavelength beyond 2400 nm due to material absorption by silica and water. We demonstrate 1580-2520 nm wavelength-tunable, high-pulse energy soliton generation using soliton self-frequency shift in a large-mode-area silica fiber pumped by a compact fiber source. Soliton pulses with pulse width of ~100 fs and pulse energy up to 73 nJ were obtained. Second harmonic generation of the solitons enables a wavelength-tunable femtosecond source from 950 nm to 1260 nm, with pulse energy up to 21 nJ. Using such energetic pulses, we demonstrate in vivo two-photon excited fluorescence imaging of vasculature and neurons in a mouse brain at wavelength beyond the tuning range of a mode-locked Ti:Sapphire lasers.

Fiber-based tunable repetition rate source for deep tissue two-photon fluorescence microscopy.

Deep tissue multiphoton imaging requires high peak power to enhance signal and low average power to prevent thermal damage. Both goals can be advantageously achieved through laser repetition rate tuning instead of simply adjusting the average power. We show that the ideal repetition rate for deep two-photon imaging in the mouse brain is between 1 and 10 MHz, and we present a fiber-based source with an arbitrarily tunable repetition rate within this range. The performance of the new source is compared to a mode-locked Ti:Sapphire (Ti:S) laser for in vivo imaging of mouse brain vasculature. At 2.5 MHz, the fiber source requires 5.1 times less average power to obtain the same signal as a standard Ti:S laser operating at 80 MHz.

Characterization and adaptive compression of a multi-soliton laser source.

Ultrashort pulse generation in the 1600 nm wavelength region is required for deep-tissue biomedical imaging. We report on the characterization and adaptive compression of a multi-soliton output spanning >300 nm from a large-mode area photonic-crystal fiber rod for two separate laser setups. Sub-30 fs pulses are generated by first compressing of each soliton individually, and then followed by coherently combining all of the pulses in the train, which are separated by hundreds of femtoseconds. Simulations of the source, together with amplitude and phase coherence measurements are provided.

Nonresonant background suppression for coherent anti-Stokes Raman scattering microscopy using a multi-wavelength time-lens source.

We demonstrate a robust, all-fiber, two-wavelength time-lens source for background-free coherent anti-Stokes Raman scattering imaging. The time-lens source generates two picosecond pulse trains simultaneously: one at 1064 nm and the other tunable between 1040 nm and 1075 nm (~400 mW for each wavelength). When synchronized to a mode-locked Ti:Sapphire laser, the two wavelengths are used to obtain on- and off-resonance coherent anti-Stokes Raman scattering images. Real-time subtraction of the nonresonant background in the coherent anti-Stokes Raman scattering image is achieved by the synchronization of the pixel clock and the time-lens source. Background-free coherent anti-Stokes Raman scattering imaging of sebaceous glands in ex vivo mouse tissue is demonstrated.

Time-lens based hyperspectral stimulated Raman scattering imaging and quantitative spectral analysis.

We demonstrate a hyperspectral stimulated Raman scattering (SRS) microscope through spectral-transformed excitation. The 1064 nm Stokes pulse was from a synchronized time-lens source, generated through time-domain phase modulation of a continuous wave (CW) laser. The tunable pump pulse was from linear spectral filtering of a femtosecond laser output with an intra-pulse spectral scanning pulse shaper. By electronically modulating the time-lens source at 2.29 MHz, hyperspectral stimulated Raman loss (SRL) images were obtained on a laser-scanning microscope. Using this microscope, DMSO in aqueous solution with a concentration down to 28 mM could be detected at 2 µs time constant. Hyperspectral SRL images of prostate cancer cells were obtained. Multivariate curve resolution analysis was further applied to decompose the SRL images into concentration maps of CH2 and CH3 bonds. This method offers exciting potential in label-free imaging of live cells using fingerprint Raman bands.

Three-photon excited fluorescence imaging of unstained tissue using a GRIN lens endoscope.

We present a compact and portable three-photon gradient index (GRIN) lens endoscope system suitable for imaging of unstained tissues, potentially deep within the body, using a GRIN lens system of 1 mm diameter and 8 cm length. The lateral and axial resolution in water is 1.0 µm and 9.5 µm, respectively. The ~200 µm diameter field of view is imaged at 2 frames/s using a fiber-based excitation source at 1040 nm. Ex vivo imaging is demonstrated with unstained mouse lung at 5.9 mW average power. These results demonstrate the feasibility of three-photon GRIN lens endoscopy for optical biopsy.

Intermodal Cerenkov radiation in a higher-order-mode fiber.

We demonstrate an intermodal Cerenkov radiation effect in a higher-order-mode (HOM) fiber with a mode crossing (i.e., two guided modes having the same propagation constant at the same wavelength). A frequency-shifted soliton in the vicinity of the mode-crossing wavelength emits a phase-matched dispersive wave in a different propagation mode. We develop a theoretical explanation for this nonlinear optical effect and demonstrate that the mode crossing in HOM fibers can be utilized to achieve simultaneous wavelength and mode conversion; the strength of this intermodal nonlinear interaction can be tuned by controlled fiber bending.

Intermodal four-wave mixing in a higher-order-mode fiber.

We demonstrate a high-efficiency intermodal four-wave-mixing process in an all-fiber system, comprising a picosecond fiber laser and a high-order-mode (HOM) fiber. Two pump photons in the LP(01) mode of the fiber can generate an anti-Stokes photon in the LP(01) mode and a Stokes photon in the LP(02) mode. The wavelength dependent mode profiles of the HOM fiber produce significant spatial overlap between the modes involved. The anti-Stokes wave at 941 nm is generated with 20% conversion efficiency with input pulse energy of 20 nJ. The guidance of the anti-Stokes and Stokes waves in the HOM fiber enhances system stability.

Higher-order-mode fiber optimized for energetic soliton propagation.

We describe the design optimization of a higher-order-mode (HOM) fiber for energetic soliton propagation at wavelengths below 1300 nm. A new HOM fiber is fabricated according to our design criteria. The HOM fiber is pumped at 1045 nm by an energetic femtosecond fiber laser. The soliton self-frequency shift process shifts the center wavelength of the soliton to 1085 nm. The soliton has a temporal duration of 216 fs and a pulse energy of 6.3 nJ. The demonstrated pulse energy is approximately six times higher than the previous record in a solid core fiber at wavelengths below 1300 nm.