Abstract
Stimulated Raman scattering (SRS) is suitable for combination with superresolution microscopy techniques such as Stimulation Depletion Emission (STED) microscopy to explore nanoscale biological processes. Whereas SRS may be employed for label-free imaging of lipids and proteins, STED microscopy allows for super-resolution of small labelled-vesicles such as exosomes, which play a key role in cell-to-cell communication. The combination of these two imaging techniques allows for more comprehensive study of biological samples under investigation. Herein, we implemented STED microscopy onto an existing custom SRS setup. Herein, the same stoke and pump beam used for SRS were employed for excitation and depletion beam on STED microscopy. The pulse widths of the picosecond lasers were independently adjusted for efficient SRS and STED imaging. The point spread function was engineered to alternate between a conventional Gaussian and Laguerre-Gaussian donut beam for sequential SRS and STED imaging, respectively. The similarity of the techniques facilitated their combination.
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