Shot-noise limited tunable dual-vibrational frequency stimulated Raman scattering microscopy.

We present a shot-noise limited SRS implementation providing a >200 mW per excitation wavelength that is optimized for addressing two molecular vibrations simultaneously. As the key to producing a 3 ps laser of different colors out of a single fs-laser (15 nm FWHM), we use ultra-steep angle-tunable optical filters to extract 2 narrow-band Stokes laser beams (1-2 nm & 1-2 ps), which are separated by 100 cm-1. The center part of the fs-laser is frequency doubled to pump an optical parametric oscillator (OPO). The temporal width of the OPO's output (1 ps) is matched to the Stokes beams and can be tuned from 650-980 nm to address simultaneously two Raman shifts separated by 100 cm-1 that are located between 500 cm-1 and 5000 cm-1. We demonstrate background-free SRS imaging of C-D labeled biological samples (bacteria and Drosophila). Furthermore, high quality virtual stimulated Raman histology imaging of a brain adenocarcinoma is shown for pixel dwell times of 16 µs.

High-repetition rate, mid-infrared, picosecond pulse generation with µJ-energies based on OPG/OPA schemes in 2-µm-pumped ZnGeP2.

We report on two different concepts to generate µJ-level mid-infrared laser pulses with sub-10 ps pulse duration via nonlinear parametric generation at high pulse repetition rates. Both schemes rely on the recent development of compact and efficient CPA-free, Ho:YLF-based 2-µm laser sources pumping the highly nonlinear crystal ZnGeP2 used for parametric amplification. The first concept comprises a simplified OPG/OPA scheme efficiently producing signal and idler radiation at fixed wavelengths of 3.0 and 6.5 µm, respectively. In the second concept, we demonstrate a wide spectral tunability of the picosecond, µJ pulses over the entire tuning range between 2.5 and 12 µm maintaining a constant bandwidth of sub-20 cm-1 by integrating a two-color pumping scheme and a spectral shaper into the setup. The presented compact and efficient mid-IR picosecond radiation sources offer a way towards low-cost mid-IR material processing and spectroscopic applications.

Background-suppressed SRS fingerprint imaging with a fully integrated system using a single optical parametric oscillator.

Stimulated Raman Scattering (SRS) imaging can be hampered by non-resonant parasitic signals that lead to imaging artifacts and eventually overwhelm the Raman signal of interest. Stimulated Raman gain opposite loss detection (SRGOLD) is a three-beam excitation scheme capable of suppressing this nonlinear background while enhancing the resonant Raman signal. We present here a compact electro-optical system for SRGOLD excitation which conveniently exploits the idler beam generated by an optical parametric oscillator (OPO). We demonstrate its successful application for background suppressed SRS imaging in the fingerprint region. This system constitutes a simple and valuable add-on for standard coherent Raman laser sources since it enables flexible excitation and background suppression in SRS imaging.

Origin and suppression of parasitic signals in Kagomé lattice hollow core fibers used for SRS microscopy and endoscopy.

Hollow core fibers are considered as promising candidates to deliver intense temporally overlapping picosecond pulses in applications such as stimulated Raman scattering (SRS) microscopy and endoscopy because of their inherent low nonlinearity compared to solid-core silica fibers. Here we demonstrate that, contrary to prior assumptions, parasitic signals are generated in Kagomé lattice hollow core fibers. We identify the origin of the parasitic signals as an interplay between the Kerr nonlinearity of air and frequency-dependent fiber losses. Importantly, we identify the special cases of experimental parameters that are free from parasitic signals, making hollow core fibers ideal candidates for noise-free SRS microscopy and endoscopy.

Real-time Raman and SRS imaging of living human macrophages reveals cell-to-cell heterogeneity and dynamics of lipid uptake.

Monitoring living cells in real-time is important in order to unravel complex dynamic processes in life sciences. In particular the dynamics of initiation and progression of degenerative diseases is intensely studied. In atherosclerosis the thickening of arterial walls is related to high lipid levels in the blood stream, which trigger the lipid uptake and formation of droplets as neutral lipid reservoirs in macrophages in the arterial wall. Unregulated lipid uptake finally results in foam cell formation, which is a hallmark of atherosclerosis. In previous studies, the uptake and storage of different fatty acids was monitored by measuring fixed cells. Commonly employed fluorescence staining protocols are often error prone because of cytotoxicity and unspecific fluorescence backgrounds. By following living cells with Raman spectroscopic imaging, lipid uptake of macrophages was studied with real-time data acquisition. Isotopic labeling using deuterated palmitic acid has been combined with spontaneous and stimulated Raman imaging to investigate the dynamic process of fatty acid storage in human macrophages for incubation times from 45 min to 37 h. Striking heterogeneity in the uptake rate and the total concentration of deuterated palmitic acid covering two orders of magnitude is detected in single as well as ensembles of cultured human macrophages. SRS signal of deuterated palmitic acid measured at the CD vibration band after incorporation into living macrophages.

Single-step sub-200 fs mid-infrared generation from an optical parametric oscillator synchronously pumped by an erbium fiber laser.

We demonstrate the single-step generation of mid-infrared femtosecond laser pulses in a AgGaSe2 optical parametric oscillator that is synchronously pumped by a 100 MHz repetition rate sub-90 fs erbium fiber laser. The tuning range of the idler beam in principle covers ∼3.5 to 17 µm, only dependent on the choice of cavity and mirror design. As an example, we experimentally demonstrate idler pulse generation from 4.8 to 6.0 µm optimized for selective vibrational resonant molecular spectroscopy. We find an oscillation threshold as low as 150 mW of pump power. At 300 mW pump power and a central wavelength of ∼5.0 µm, we achieve an average infrared power of up to 17.5 mW, with a photon conversion efficiency of ∼18%. A pulse duration of ∼180 fs is determined from a nonlinear cross-correlation with residual pump light. The single-step nonlinear conversion leads to a high power stability with <1% average power drift at <0.5% rms noise over 1 h.

Difference-frequency generation of ultrashort pulses in the mid-IR using Yb-fiber pump systems and AgGaSe(2).

We employ AgGaSe(2) for difference-frequency generation between signal and idler of synchronously-pumped picosecond / femtosecond OPOs at 80 / 53 MHz. Continuous tuning in the picosecond regime is achieved from 5 to 18 µm with average power of 140 mW at 6 µm. In the femtosecond regime the tunability extends from 5 to 17 µm with average power of 69 mW at 6 µm. Maximum single pulse energies of >1 nJ in both cases represent the highest values at such high repetition rates.

Difference-frequency generation of fs and ps mid-IR pulses in LiInSe(2) based on Yb-fiber laser pump sources.

Difference-frequency generation between signal and idler of Yb-fiber laser synchronously pumped femtosecond and picosecond optical parametric oscillators (SPOPOs) operating at 53 and 80 MHz, respectively, is investigated in the wide bandgap chalcogenide crystal LiInSe(2). Single pulse energies in excess of 1 nJ are demonstrated, and the continuous tuning extends from 5 to 12 µm.