Low-noise, fast-tuning light source for coherent Raman microscopy and stimulated Raman microscopy
▍Background principle
Optical imaging technology has become a widely used microscopic imaging technology with the characteristics of non-contact measurement, no damage to the sample, high spatiotemporal resolution and high detection sensitivity. The fluorescent labeled imaging method has the advantages of high acquisition sensitivity and a wide variety of fluorescent labels, and is widely used in the biomedical field. However, its application is limited by its interference and photobleaching. Based on infrared spectroscopy and Raman spectroscopy microscopy technology to detect the natural vibration frequency of sample molecules, biological samples do not need to be pre-labeled, and directly use the characteristic spectral signal as imaging contrast, which has the advantages of molecular feature selectivity, and it has been widely used in biomedical microscopy, such as live cell, tissue or DNA imaging. However, the molecular spectral signal intensity is weak, which limits the detection sensitivity, laser power and data acquisition. Coherent Anti-Stokes Raman Scattering (CARS), which is also based on molecular vibrational energy levels, is a third-order nonlinear optical process, which produces a strong stimulated resonance signal and a certain directionality, which makes the CARS signal collection more efficient.
▍ Appliance device
Although CARS are very weak, they are spectrally distinguishable. But for stimulated Raman scattering, small changes in the pump light or Stokes beam must be measured. Therefore, the latter needs to be detected based on lock-in amplification techniques, which means that for SRS, ultra-low laser noise intensity is required, ideally below the shot noise limit. At the same time, laser scanning imaging requires good long-term average power stability and reproducibility, as well as fast tuning while maintaining its spatiotemporal properties.
The vast majority of CRS work is based on picoseconds, pumped optical parametric oscillators (OPOs), and timing-synchronized Titanium-sapphire laser systems. Stuttgart Instruments offers wideband, fast-tuning, and low-noise CRS light sources with excellent stability using a ytterbium-doped solid-state laser-pumped fiber feedback OPO system.
The system uses a ytterbium-doped solid-state laser as the pump light source, generating a central wavelength of 1043nm, an average power of 6W, about 390fs (marked as 320fs in the figure), and a repetition rate of 75MHz. Among them, 2.5W is used to pump OPO to generate a 1380-2000nm near-infrared laser, which is narrowed on the spectrum through an air gap etalon to produce a Stokes beam with a pulse width of 600mW and 1.3ps. The frequency doubling is then generated by PPLN as the Raman pump light. In order to ensure the synchronization of pulse timing, a delay line is added to the Stokes beam path for adjustment.
▍Characteristics of light source
The near-infrared laser generated by OPO can be tuned between 1.38um and 2.02um, with an output linewidth of 9-20nm and a pulse width of 200-300fs, and the Stokes optical power after staltalon is 600mW@1043.1nm and 0.71nm, corresponding to a CRS measurement resolution of 6.5cm-1.
It produces 750nm~940nm pump light, and the output is close to the Fourier limit in the whole tuning range, with a typical pulse duration of 1.1ps. Stokes reaches the shot noise limit at 150 KHz, the OPO and pump light are slightly higher, but the shot noise limit can be reached above 1 MHz, and a low-noise CRS spectrum can be obtained.