Raman microscopy is a powerful tool for analytical imaging. The wavelength shift of Raman scattering corresponds to molecular vibrational energy. Therefore, we can access rich chemical information, such as distribution, concentration, and chemical environment of sample molecules. Despite these strengths of Raman microscopy, the spatial resolution has been a limiting factor for many practical applications. In this study, we developed a large-area, high-resolution Raman microscope by utilizing structured illumination microscopy (SIM) to overcome the spatial resolution limit.
A structured line-illumination (SLI) Raman microscope was constructed. The structured illumination is introduced along the line direction by the interference of two line-shaped beams. In SIM, the spatial frequency mixing between structured illumination and Raman scattering from the sample allows access to the high spatial frequency information beyond the conventional cut-off. As a result, the FWHM of 40-nm fluorescence particle images showed a clear resolution enhancement in the line direction: 366 nm in LI and 199 nm in SLI microscope. Using the developed microscope, we successfully demonstrated high-resolution Raman imaging of various kinds of specimens, such as few-layer graphene, graphite, mouse brain tissue, and polymer nanoparticles.
The high resolution Raman images showed the capability to extract original spectral features from the mixed Raman spectra of a multi-component sample because of the enhanced spatial resolution, which is advantageous in observing complex spectral features. The Raman microscopy technique reported here enables us to see the detailed chemical structures of chemical, biological, and medical samples with a spatial resolution smaller than 200 nm.
Kozue Watanabe, Almar F. Palonpon, Nicholas I. Smith, Liang-da Chiu, Atsushi Kasai, Hitoshi Hashimoto, Satoshi Kawata, and Katsumasa Fujita, "Structured illumination microscopy for vibrational molecular imaging
(Conference Presentation)," Proc. SPIE 9925, Nanoimaging and Nanospectroscopy IV, 992504 (Presented at SPIE Nanoscience + Engineering: August 28, 2016; Published: 3 November 2016); https://doi.org/10.1117/12.2239007.5161498093001.
Conference Presentations are recordings of oral presentations given at SPIE conferences and published as part of the proceedings. They include the speaker's narration with video of the slides and animations. Most include full-text papers. Interactive, searchable transcripts and closed captioning are now available for most presentations.
Search our growing collection of more than 22,000 conference presentations, including many plenaries and keynotes.