Abstract
Label-free vibrational imaging by stimulated Raman scattering (SRS) provides unprecedented insight into real-time chemical distributions. Specifically, SRS in the fingerprint region (400–1800 cm−1) can resolve multiple chemicals in a complex bio-environment. However, due to the intrinsic weak Raman cross-sections and the lack of ultrafast spectral acquisition schemes with high spectral fidelity, SRS in the fingerprint region is not viable for studying living cells or large-scale tissue samples. Here, we report a fingerprint spectroscopic SRS platform that acquires a distortion-free SRS spectrum at 10 cm−1 spectral resolution within 20 µs using a polygon scanner. Meanwhile, we significantly improve the signal-to-noise ratio by employing a spatial-spectral residual learning network, reaching a level comparable to that with 100 times integration. Collectively, our system enables high-speed vibrational spectroscopic imaging of multiple biomolecules in samples ranging from a single live microbe to a tissue slice.
Highlights
Label-free vibrational imaging by stimulated Raman scattering (SRS) provides unprecedented insight into real-time chemical distributions
Stimulated Raman scattering (SRS) microscopy is a highspeed vibrational imaging modality that produces chemical maps in dynamic living systems based on intrinsic molecular vibrations[1,2,3,4,5]
The Stokes beam is sent to a 55kHz polygon scanner and subsequently scanned to a blazed grating at Littrow configuration, which acts as a wedge to introduce a continuous-changing path difference between the pump and the retroreflected Stokes beam
Summary
Label-free vibrational imaging by stimulated Raman scattering (SRS) provides unprecedented insight into real-time chemical distributions. An existing high-speed spectral focusing scheme[21] by an edge-reflected resonant mirror has only 2-ps delay range, which limits the degree of chirping and leads to 28 cm−1 spectral resolution that is not sufficient for the fingerprint region. Another galvo-based delay-line scanning setup can achieve a large tuning range and high linearity[22], but the scanning speed is limited to 1 kHz
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