We present a femtosecond laser system at 920 nm delivering ultrashort pulses via a hollow-core photonic bandgap fiber (HC-PBGF). The laser system is designed to simplify two-photon microscopy applications and can be used for miniaturized two-photon microscopes. While previously presented solutions have been tailored to a specific length and dispersion coefficient of the HC-PBGF, we now show a compact and flexible scheme for dispersion compensation which is compatible with a wide range of fiber types and lengths.
In addition, this new approach fully maintains the capability of software-controlled dispersion compensation in the range from 0 to -40,000 fs2 after the pulse delivery fiber. Hence, the dispersion of common two-photon microscopes can be pre-compensated in order to obtain compressed pulses at the sample plane. Our newly developed system displays excellent long-term fiber coupling stability under varying environmental conditions. It is capable of polarization-preserving femtosecond pulse delivery at 920 nm and reaches Watt-level power after the delivery fiber, making it suitable for in-vivo brain imaging of GCaMP in mouse models.
We present time resolved measurements on low dimensional nanomaterials like individual (6,4) single-walled carbon nanotubes and monolayers of MoSe2 via transient interferometric scattering (TiSCAT) microscopy. For this a novel fiber laser system was developed comprising a tunable probe arm and low noise performance. The sensitivity of the measurement is demonstrated for very low excitation powers to prevent photodamage of the sample. Signal variations close to the shot-noise limit can be resolved even with low excitation powers in the order of 1 μW. In combination with the tunability of the laser system the absorption spectrum of a single SWCNT was determined.
We will present our latest innovations about ultrafast fiber lasers and show how multiphoton microscopy can benefit from these developments. Wavelengths around 900 nm and pulse durations as short as 100 fs remain a challenge for fiber lasers. Here we present a two-color femtosecond fiber laser system with synchronous outputs. One arm emits pulses at a central wavelength of 780 nm and the novel second laser arm is continuously tunable in its central wavelength between 810 nm and 950 nm. This allows the independent excitation of NADH and FAD and therefore enables optical metabolism and oxygen imaging of cells via FLIM and PLIM measurements.
We describe a metabolic-imaging system based on simultaneous recording of lifetime images of NAD(P)H and FAD. The system uses two-photon excitation by a dual-wavelength femtosecond fibre laser. The two wavelengths of the laser, 780 nm and 880 nm, are multiplexed synchronously with the frames or the lines of the scan. The recording system uses two parallel TCSPC FLIM channels, detecting from 420 to 475 nm and 480 to 600 nm. By using the multiplexing functions of the TCSPC modules, separate images for NAD(P)H and FAD are recorded. A third image is obtained for the SHG of the 880 nm laser wavelength. Data analysis delivers images of the amplitude-weighted lifetime, tm, the component lifetimes, t1 and t2, the amplitudes of the components, a1 and a2, the amplitude ratio, a1/a2, and the fluorescence-lifetime redox ratio (FLIRR), a2nadh/a1fad. We demonstrate the performance of the system for metabolic imaging of mammalian skin.
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