Katsumasa Fujita

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We demonstrate dynamic imaging of molecular distribution in unstained living cells using Raman scattering. By combining slit-scanning detection and optimizing the excitation wavelength, we imaged the dynamic molecular distributions of cytochrome c, protein beta sheets, and lipids in unstained HeLa cells with a temporal resolution of 3 minutes. We found that(More)
Label-free imaging is desirable for elucidating morphological and biochemical changes of heart tissue in vivo. Spontaneous Raman microscopy (SRM) provides high chemical contrast without labeling, but presents disadvantage in acquiring images due to low sensitivity and consequent long imaging time. Here, we report a novel technique for label-free imaging of(More)
Raman microscopy is a promising technology for visualizing the distribution of molecules in cells. A challenge for live-cell imaging using Raman microscopy has been long imaging times owing to the weak Raman signal. Here we present a protocol for constructing and using a Raman microscope equipped with both a slit-scanning excitation and detection system and(More)
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(More)
Alkyne has a unique Raman band that does not overlap with Raman scattering from any endogenous molecule in live cells. Here, we show that alkyne-tag Raman imaging (ATRI) is a promising approach for visualizing nonimmobilized small molecules in live cells. An examination of structure-Raman shift/intensity relationships revealed that alkynes conjugated to an(More)
A recent key requirement in life sciences is the observation of biological processes in their natural in vivo context. However, imaging techniques that allow fast imaging with higher resolution in 3D thick specimens are still limited. Spinning disk confocal microscopy using a Yokogawa Confocal Scanner Unit, which offers high-speed multipoint confocal live(More)
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(More)
We propose a multifocus scanning technique in second-harmonic-generation (SHG) microscopy in which the SHG detection efficiency and the image acquisition rate are improved by several tens of times compared with typical single-focus scanning techniques. Because of a microlens array scanner, the laser beam of a mode-locked Ti:sapphire laser is split into ~100(More)
Raman scattering microscopy provides information about the distribution and chemical state of molecules in live cells without any labeling or modification. In recent years, the imaging speed of Raman microscopy has improved greatly owing to the development of instruments that can perform parallel acquisition of Raman spectra from multiple points. In this(More)
Ischemic insult to the heart produces myocyte Ca2+ ([Ca2+]i) overload. However, little is known about spatiotemporal changes in [Ca2+]i within the ischemic heart in situ at the cellular level. Using real-time confocal microscopy, we successfully visualized [Ca2+]i dynamics at the border zone on the subepicardial myocardium of the heart 2 h after coronary(More)