Abstract
In the last couple of decades, the spatial resolution in optical microscopy has increased to unprecedented levels by exploiting the fluorescence properties of the probe. At about the same time, Raman imaging techniques have emerged as a way to image inherent chemical information in a sample without using fluorescent probes. However, in many applications, the achievable resolution is limited to about half the wavelength of excitation light. Here we report the use of structured illumination to increase the spatial resolution of label-free spontaneous Raman microscopy, generating highly detailed spatial contrast from the ensemble of molecular information in the sample. Using structured line illumination in slit-scanning Raman microscopy, we demonstrate a marked improvement in spatial resolution and show the applicability to a range of samples, including both biological and inorganic chemical component mapping. This technique is expected to contribute towards greater understanding of chemical component distributions in organic and inorganic materials.
Highlights
In the last couple of decades, the spatial resolution in optical microscopy has increased to unprecedented levels by exploiting the fluorescence properties of the probe
High spectral resolution and relatively high imaging speed are maintained in structured-lineillumination (SLI) Raman microscopy because of the parallel detection of Raman scattering through the spectrophotometer slit
Raman bands is simultaneously available, making it a more potentially useful tool for diagnosis. These results show that with different chemical components in closer proximity than the resolution capability of standard Raman imaging, the improvement of the spatial resolution contributes to a more detailed spectroscopic analysis using Raman scattering with spectral discrimination of the distributed components. In these experiments, the use of structured line illumination in Raman microscopy shows spatial resolution improvements and the increased capability to discriminate component spectra, which is advantageous for the analysis of multi-component samples whose Raman spectra are complicated
Summary
In the last couple of decades, the spatial resolution in optical microscopy has increased to unprecedented levels by exploiting the fluorescence properties of the probe. The approaches toward improvement of the spatial resolution in optical microscopy have been successful in the last couple of decades In these achievements, the excitation and emission properties of fluorescence probes play a key role to overcome the classical spatial resolution limitation[1]. Structured illumination microscopy (SIM) illuminates a sample by using an illumination with a grid pattern producing the moireeffect to make small structure information transferable through the imaging optics, resulting in a doubling of the spatial resolution compared with the classical limit[8,9] These approaches are usually grounded in modification of a fluorescence-based imaging method. High spectral resolution and relatively high imaging speed are maintained in structured-lineillumination (SLI) Raman microscopy because of the parallel detection of Raman scattering through the spectrophotometer slit Without both of these features, the possible applications are restricted.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
