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Related Concept Videos

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy09:57

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

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We present a protocol to acquire chemical images with broadband stimulated Raman scattering (SRS) microscopy. Based on an SRS microscope that operates with differential multichannel-lock-in detection, the protocol describes the sample preparation, adjustment of the SRS apparatus, and chemometrics to disentangle different constituents of chemically heterogeneous...
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Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging09:46

Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging

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This paper directly compares the resolution, sensitivity, and imaging contrasts of stimulated Raman scattering (SRS) and coherent anti-Stokes Raman scattering (CARS) integrated into the same microscope platform. The results show that CARS has a better spatial resolution, SRS gives better contrasts and spectral resolution, and both methods have similar sensitivity.
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Two-Color Stimulated Raman Scattering Imaging of Mouse Brain Tissue07:16

Two-Color Stimulated Raman Scattering Imaging of Mouse Brain Tissue

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Source: Espinoza, R., et al. Real-Time, Two-Color Stimulated Raman Scattering Imaging of Mouse Brain for Tissue Diagnosis. J. Vis. Exp. (2022). The video demonstrates two-color stimulated Raman scattering (SRS) imaging of mouse brain tissue sections. Two synchronized laser beams are directed onto the tissue section to excite molecular vibrations, resulting in a relative intensity change in the beams. Lipids and proteins in the tissue are detected using distinct frequency...
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Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy10:59

Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy

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Stimulated Raman scattering (SRS) microscopy allows selective, label-free imaging of specific chemical moieties and it has been effectively employed to image lipid molecules in vivo. Here, we provide a brief introduction to the principle of SRS microscopy and describe methods for its use in imaging lipid storage in Caenorhabditis...
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Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering09:13

Implementation of a Nonlinear Microscope Based on Stimulated Raman Scattering

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In this manuscript, the implementation of a stimulated Raman scattering (SRS) microscope, obtained by the integration of an SRS experimental set-up with a laser scanning microscope, is described. The SRS microscope is based on two femtosecond (fs) laser sources, a Ti-Sapphire (Ti:Sa) and synchronized optical parametric oscillator...
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In vivo Imaging of Biological Tissues with Combined Two-Photon Fluorescence and Stimulated Raman Scattering Microscopy09:06

In vivo Imaging of Biological Tissues with Combined Two-Photon Fluorescence and Stimulated Raman Scattering Microscopy

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Stimulated Raman scattering (SRS) microscopy allows label-free imaging of biomolecules based on their intrinsic vibration of specific chemical bonds. In this protocol, the instrumental setup of an integrated SRS and two-photon fluorescence microscope is described to visualize cellular structures in the spinal cord of live...
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Related Experiment Video

Updated: Jan 19, 2026

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy
09:57

Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

Published on: July 25, 2022

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Orthogonal beam ballistic backscatter stimulated Raman microscopy.

Robert D Frankel

    Optics Express
    |September 13, 2019
    PubMed
    Summary

    Significant dipole-like ballistic backscatter occurs in stimulated Raman microscopes when the axial gain length is less than 40% of the emission wavelength. This enables rapid, sub-wavelength 3D imaging of label-free Raman contrast for pathology and metabolites.

    Area of Science:

    • Optics and Photonics
    • Microscopy
    • Biomedical Imaging

    Background:

    • Stimulated Raman scattering (SRS) microscopy offers label-free imaging capabilities.
    • Achieving sub-wavelength resolution and high-speed 3D imaging in SRS microscopy remains a challenge.
    • Understanding backscatter phenomena is crucial for optimizing SRS microscope design.

    Purpose of the Study:

    • To analyze a scanning SRS microscope configuration designed to induce significant dipole-like ballistic backscatter.
    • To investigate the impact of pump beam characteristics (focus and polarization) on backscattered signals.
    • To evaluate the potential for sub-wavelength resolution 3D imaging of biological samples and metabolites.

    Main Methods:

    • Utilized a scanning SRS microscope with orthogonal water-dipping objectives satisfying the criterion for ballistic backscatter.

    More Related Videos

    Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
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    Published on: April 28, 2022

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    Two-Color Stimulated Raman Scattering Imaging of Mouse Brain Tissue
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    Two-Color Stimulated Raman Scattering Imaging of Mouse Brain Tissue

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    Related Experiment Videos

    Last Updated: Jan 19, 2026

    Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy
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    Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy

    Published on: July 25, 2022

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    Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
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    Two-Color Stimulated Raman Scattering Imaging of Mouse Brain Tissue
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  • Employed Gaussian spot and droplet Bessel beams for pump focus, analyzing their impact on focal regions.
  • Investigated radial and linearly polarized pump beams to enable transverse signal detection.
  • Identified low-level Mie backscatter as the primary photon noise source.

    • Main Results:

    • Confirmed significant dipole-like ballistic backscatter under specific axial gain length conditions (<40% of emission wavelength).
    • Demonstrated that droplet Bessel beams minimize secondary lobes and create multiple focal regions.
    • Showed that radial and linear polarization enable backscattered signals along transverse axes.
    • Identified Mie backscatter as the dominant noise source.

    Conclusions:

    • The analyzed SRS microscope configuration facilitates rapid, sub-wavelength resolution 3D imaging.
    • This technique is suitable for label-free imaging of in-vivo pathology.
    • The method can image physiological concentrations of Raman-labeled metabolites and drugs.