Katsumasa Fujita

We demonstrate the use of saturated excitation in confocal fluorescence microscopy to improve the spatial resolution. In the proposed technique, we modulate the excitation intensity temporally and detect the harmonic modulation of the fluorescence signal which is caused by the saturated excitation in the center of the laser focus. Theoretical and experimental investigations show that the demodulated fluorescence signal is nonlinearly proportional to the excitation intensity and contributes to im

We perform time-resolved observation of living cells with gold nanoparticles using surface-enhanced Raman scattering (SERS). The position and SERS spectra of 50-nm gold nanoparticles are simultaneously observed by slit-scanning Raman microscopy with high spatial and temporal resolution. From the SERS observation, we confirm the attachment of the particles on the cell surface and the entry into the cell with the subsequent generation of SERS signals from nearby molecules. We also confirm that the

Intracellular pH is one of the key factors for understanding various biological processes in biological cells. Plasmonic gold and silver nanoparticles (NPs) have been extensively studied for surface-enhanced Raman scattering (SERS) applications for pH sensing as a local pH probe in a living cell. However, the SERS performance of NPs depends on material, size, and shape, which can be controlled by chemical synthesis. Here, we synthesized 18 types of gold and silver NPs with different morphologies

Visualizing live-cell uptake of small-molecule drugs is paramount for drug development and pharmaceutical sciences. Bioorthogonal imaging with click chemistry has made significant contributions to the field, visualizing small molecules in cells. Furthermore, recent developments in Raman microscopy, including stimulated Raman scattering (SRS) microscopy, have realized direct visualization of alkyne-tagged small-molecule drugs in live cells. However, Raman and SRS microscopy still suffer from limi

Optical signals based on Raman scattering, coherent anti-Stokes Raman scattering (CARS), and harmonic generation can be used to image biological molecules in living cells without labeling. Both Raman scattering and CARS signals can be used to detect frequencies of molecular vibrations and to obtain the molecular distributions in samples. Second-harmonic optical signals can also be generated in structured arrays of noncentrosymmetric molecules and can be used to detect structured aggregates of pr

Metal nanoparticles have recently emerged as ubiquitous surface-enhanced Raman scattering (SERS) agents for nano-imaging and nano-analysis. These applications make use of the unique optical properties of metal nanoparticles to enhance the efficiency of Raman scattering, whereas the small size of the nanoparticles localizes the enhanced Raman scattering at the nanoscale. In this perspective, we review the recent progress in SERS nano-imaging and nano-spectroscopy using metal nanoparticles applied

Mitochondrial activity is a widely used criterion to judge the metabolic condition of a living specimen. Numerous methods have been developed for related analyses, including the detection of O2 consumption, trans-membrane potential, and ATP production. In this study, we demonstrate that the redox state of cytochromes can serve as a sensitive mitochondrial activity indicator in glutamate-stressed neuronal cells. Mitochondrial dysfunction was detected by Raman imaging as early as 30 min after glut