A Spectral Resolution study in Femtosecond Stimulated Raman Scattering Microscopy

C Sibilia,G Pauliat,Luigi Sirleto,Maria Antonietta Ferrara,Rajeev Ranjan,A Belardini

doi:10.1051/epjconf/202125511009

C Sibilia, G Pauliat + Show 4 more

Open Access

https://doi.org/10.1051/epjconf/202125511009

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Abstract

The simultaneous mapping and the specificity of different chemical species are desirable in several biological and biomedical applications. The stimulated Raman Scattering technique is a proven and well-established label-free method to map the distributions of individual species in a multi-component-based system due to the linear dependence of signals on concentration and its chemical selectivity. In this framework, spectral resolution, i.e., the ability to distinguish closely lying resonances, plays a fundamental role. Here in this work, the cross-correlation of Ti:Sa & OPO femtosecond laser beams in a stimulated Raman scattering microscope is measured. The separation between protein and lipid bands in the C-H region is important for biochemical research and is successfully classified.

Highlights

Over the past ten years, stimulated Raman scattering (SRS) has been investigated in nanophotonic [1,2,3,4] as well as in biophotonics as an analytical, label-free, noninvasive technique with unique cellular and tissue imaging capabilities [5,6,7,8]
In this paper, taking advantage of the small chirping introduced by propagating the beams through dispersive materials already present in the SRS microscope setup, we demonstrate that we can be distinguished protein and lipid bands in the C-H region
We have successfully described the design and execution of femtosecond-based SRS microscopy, which is able to cover the C-H region (>2,500 cm-1) [16,17,18,19,20,21,22]

Summary

Introduction

Over the past ten years, stimulated Raman scattering (SRS) has been investigated in nanophotonic [1,2,3,4] as well as in biophotonics as an analytical, label-free, noninvasive technique with unique cellular and tissue imaging capabilities [5,6,7,8]. Lipids and proteins can be investigated in this region, taking advantage of CH2, near 2845 cm-1, and CH3, near 2930 cm-1, Raman bands, respectively.

Results

Conclusion