Time-lens based hyperspectral stimulated Raman scattering imaging and quantitative spectral analysis

Abstract

We demonstrate a hyperspectral stimulated Raman scattering (SRS) microscope through spectral-transformed excitation. The 1064 nm Stokes pulse was from a synchronized time-lens source, generated through time-domain phase modulation of a continuous wave (CW) laser. The tunable pump pulse was from linear spectral filtering of a femtosecond laser output with an intra-pulse spectral scanning pulse shaper. By electronically modulating the time-lens source at 2.29 MHz, hyperspectral stimulated Raman loss (SRL) images were obtained on a laser-scanning microscope. Using this microscope, DMSO in aqueous solution with a concentration down to 28 mM could be detected at 2 μs time constant. Hyperspectral SRL images of prostate cancer cells were obtained. Multivariate curve resolution analysis was further applied to decompose the SRL images into concentration maps of CH₂ and CH₃ bonds. This method offers exciting potential in label-free imaging of live cells using fingerprint Raman bands.

Keywords: hyperspectral; multivariate curve resolution; nonlinear optical microscopy; pulse shaper; stimulated Raman scattering; time-lens.