Stimulated Raman scattering simulation for imaging optimization

Abstract Two simulation programs of a stimulated Raman scattering microscopy (SRS) imaging system with lock-in amplifier (LIA) detection were developed. SRS is an technique based on the vibrational cross-section as contrast mechanism and enables fast, label-free imaging. Most implementations are LIA modulated signal. However, building operating such set-ups still poses challenge when selecting parameter settings for optimized acquisition speed or image quality. Moreover, type sample, e.g. sparse sample vs. densely packed required resolution well laser powers affect choice. A program was used to find these optimal parameters. The focal spot diameters individual lasers (pump Stokes) estimate effective signal (optical) spatial resolution. By calibrating noise propagation through known molecule, we estimated input LIA. We low pass filter model simulate behavior in order parameters (i.e. time constant). Optimization done either quality (expressed ratio) time. targeted object size first determined measure output consisted parameters, pixel dwell ratio. In second evaluated same principles setting simulation, i.e. signals propagated detection. simulated images compared experimental polystyrene beads. Finally, software multiplexed this study modeled six-channel frequency-encoded demodulated six channels. inter-channel crosstalk function chosen which multiplex also needs be considered. These optimize ratio, speed, will useful setup, designing novel setups.