Stimulated Raman scattering (SRS) enables fast, high resolution imaging of chemical constituents important to biological structures and functional processes. While this technology has shown remarkable potential, it is currently limited to point scanning and can only probe a few Raman bands at a time. In this work we take a fundamentally different approach to detecting the small nonlinear signals based on dispersion effects that accompany the loss/gain processes in SRS. We use a modified pump-probe system (pulses with duration of ~0.5 ps and 75 fs, respectively) with interferometric detection in the Fourier-domain to demonstrate that the dispersive measurements are more robust to noise (e.g., laser noise) compared to conventional amplitude measurements, which in turn permits facile spectral and spatial multiplexing.
Results show that it is possible to assess a broadband dispersion spectrum (currently limited to ~400 cm-1) with a single laser shot or spectrometer acquisition (20-50 µs). For molecular imaging with broadband spectral information, we achieve spatial pixel rates of 2.5 kHz, and will discuss how this can be further improved to 20-50 kHz. We also combine SRS with optical coherence tomography (OCT) (molecular and structural information are rendered from the same data), which enables axial multiplexing by coherence gating and paves the way for volumetric biochemical imaging. The approach is tested on a thin water-and-oil phantom, a thick scattering polystyrene bead phantom, and thick freshly excised human adipose tissue. Finally, we will outline other opportunities for spatial multiplexing using wide-field holography and spectroscopic-OCT, which would massively parallelize the spatial and spectral information. The combination of dispersion-based SRS and phase imaging has the potential to enable faster wide-area and volumetric molecular imaging. Such methods would be valuable in a clinical setting for many applications.
Francisco E. Robles, Martin C. Fischer, and Warren S. Warren, "Dispersion-based stimulated Raman scattering spectroscopy, holography, and optical coherence tomography
(Conference Presentation)," Proc. SPIE 9712, Multiphoton Microscopy in the Biomedical Sciences XVI, 97120J (Presented at SPIE BiOS: February 14, 2016; Published: 27 April 2016); https://doi.org/10.1117/12.2212875.4848767232001.
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