Direct imaging of Stimulated Raman scattering : 3D spatial control and spatial generation

Stimulated Raman scattering (SRS) is a powerful imaging technique that has become popular during the last decades for its ability to image species specific in a sample with high accuracy. The purpose of this thesis is twofold. Firstly, to demonstrate 3D spatial control of where in the sample SRS is generated. Secondly, the spatial behavior of the SRS generation is investigated by experiments and simulations. SRS is a nonlinear scattering phenomenon that is produced when a sample is illuminated with two laser beams, called Stokes and pump beams, whose frequency difference corresponds to a molecular vibration caused by inelastic scattering of an incoming photon. The Stokes beam will stimulate the scattering of the pump beam photons, which leads to an intensity gain in the Stokes beam and an intensity loss in the pump beam. Imaging of SRS is usually performed by point scanning a sample in a laser scanning microscope by the two laser beams. Thereafter, the image is constructed pixel by pixel by detecting either the gain or the loss. Our aim is to perform direct field of view SRS imaging. Two experimental setups are presented in this thesis, one for the 3D spatial control of SRS and one for the investigation of the spatial generation of SRS. The working principle of imaging is the same in both setups. A cylindrical sample volume was illuminated with the Stokes beam and the SRS was generated by focusing the pump beam into this volume. The diameter of the illuminated cylinder was around 10 mm. The two beams were combined before the sample using a dichroic mirror and after the sample the pump beam was removed by a second dichroic mirror. The Stokes light was then image onto a camera providing a field of view of around 9.4 mm by 7.94 mm. A phase spatial light modulator (SLM) was used to control the shape and position of the pump beam in three dimensions (3D) in the illuminated volume. The results show that the SLM allowed for control of the position and shape of the generated SRS signal. In the second experimental setup the pump beam was focused into the sample by a lens and the spatial generation of the SRS was investigated. A second dichroic mirror blocking the pump beam was inserted into the sample at different interaction lengths to study the resulting SRS signal. Further, the pump intensity was varied to study the effect on the physical width of the SRS signal. The experimental results were compared to computer simulations. The simulations were based on diffraction theory for the beam propagation and the interaction between the light beams and the material was modeled with a phase modulation due to the induced Kerr effect caused by high pump intensity. The results shows that most of the SRS generation takes place close to the focus of the pump beam