Abstract
We report a shot noise limited high-speed stimulated Raman microscopy platform allowing to acquire molecular vibrational spectra over 200 cm-1 in 12 µs at a scan rate of 40kHz. Using spectral focusing together with optimized acousto-optics programmable dispersive filters, the designed low noise imaging platform performs chemical imaging of dynamical processes such as Mannitol crystal hydration and reaches a signal to noise ratio sufficient to perform label free histological imaging on frozen human colon tissue slides.
Highlights
Stimulated Raman scattering (SRS) [1] imaging has gain tremendous interest over the last decade due to its ability to perform label free chemical imaging in biological sample [2,3]
The noise brought by the acousto-optic programmable dispersive filters (AOPDF) was not studied, the sensitivity was not showcased on biological tissues, and the increased acquisition rate was not applied to the study of chemically dynamic behaviors, while those two aspects are essential for applications
Fast label free imaging of biopsy sections is a major application of coherent Raman imaging that can potentially revolutionize the field of histology by increasing diagnosis speed, lowering hospital costs, and improving patient care [25]
Summary
Stimulated Raman scattering (SRS) [1] imaging has gain tremendous interest over the last decade due to its ability to perform label free chemical imaging in biological sample [2,3]. In SRS microscopy two laser pulses, a pump at frequency ωp and a Stokes at frequency ωs, are focused on a sample to generate an image by point scanning. The fastest approaches to acquire a full spectrum [11,12] use the SRS spectral focusing scheme that allow to retrieve spectral resolution by adding dispersion on the femtosecond pulses [17,18]. By characterizing the noise in our system and engineering the optimal laser pulses, we have designed an SRS imaging platform combining spectral focusing together with a faster AOPDF delay line to surpass the acquisition speed and sensitivity. We demonstrate optimized speed and sensitivity by targeting two major applications that are dynamic chemical imaging and label-free histology on human tissue samples
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