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High speed imaging is pre-requisite for monitoring of dynamic processes in biological events. Here we report the development of a unique spatial light-modulated stimulated Raman scattering (SLM-SRS) microscopy that tailors the broadband excitation beam with sparse-sampling masks designed for rapid multiplexed vibrational imaging to monitor real-time cancer treatment effects and in vivo transport of drug solvent. Methods: We design an optimal mask pattern that enables selection of predominant windows in SRS spectrum for collective excitation at the highest possible peak power, thus providing an improved signal-to-noise ratio (SNR) without compromise of chemical specificity. The mask pattern generated is applied to the broad excitation beam using a flexible spatial light modulator. The SLM module further offers complementary function whereby rapid scanning of SRS spectrum can be facilitated prior to the mask generation, thereby making the SLM-SRS system a stand-alone imaging platform. Results: We demonstrate that SLM-SRS microscopy permits rapid multiplexed SRS imaging of polystyrene and polymethyl methacrylate beads in Brownian motion in dimethyl sulfoxide (DMSO) at 70 ms intervals without motion artiacts. We further apply SLM-SRS to monitor the therapeautic effect of mild alkaline solution on cancer cells, which shows immediate apoptotic response. Finally, we visualize in vivo penetration of DMSO into the plant tissue and evaluate acute toxicity of DMSO on cellulose and proteins within the tissue. Conclusion: We develop novel SLM-SRS microscopy and affirm its broad applicability for rapid monitoring of dynamic biological processes at the subcellular and molecular level.

Spatial light-modulated stimulated Raman scattering (SLM-SRS) microscopy for rapid multiplexed vibrational imaging