Abstract:In vivo two-photon microscopy is an advantageous technique for observing the mouse brain at high resolution. In this study, we developed a two-photon microscopy method that uses a 1064-nm gain-switched laser diode-based light source with average power above 4 W, pulse width of 7.5-picosecond, repetition rate of 10-MHz, and a high-sensitivity photomultiplier tube. Using this newly developed two-photon microscope for in vivo imaging, we were able to successfully image hippocampal neurons in the dentate gyrus and obtain panoramic views of CA1 pyramidal neurons and cerebral cortex, regardless of age of the mouse. Fine dendrites in hippocampal CA1 could be imaged with a high peak-signal-tobackground ratio that could not be achieved by titanium sapphire laser excitation. Finally, our system achieved multicolor imaging with neurons and blood vessels in the hippocampal region in vivo. These results indicate that our two-photon microscopy system is suitable for investigations of various neural functions, including the morphological changes undergone by neurons during physiological phenomena.
In this study, we investigated the picosecond optical pulse generation from a 1064-nm distributed feedback laser diode under strong gain switching. The spectrum of the generated optical pulses was manipulated in two different ways: (i) by extracting the short-wavelength components of the optical pulse spectrum and (ii) by compensating for spectral chirping in the extracted mid-spectral region. Both of these methods shortened the optical pulse duration to approximately 7 ps. These optical pulses were amplified to over 20-kW peak power for two-photon microscopy. We obtained clear two-photon images of neurons in a fixed brain slice of H-line mouse expressing enhanced yellow fluorescent protein. Furthermore, a successful experiment was also confirmed for in vivo deep region H-line mouse brain neuron imaging.
Sputtering enables uniform and clean deposition over a large area, which is an issue with exfoliation and chemi-cal vapor deposition methods. On the other hand, the process of physical vapor deposition (PVD) film formationhas not yet been clarified. We prepared several samples from the sub-monolayer region, and performed Ra-man spectroscopy, X-ray photon spectroscopy and high-angle annular dark-field scanning transmission electronmicroscopy. From these results, the internal stresses inherent to PVD films, the bonding states specific to sub-monolayers, and the unique film structure and the grain formation process of PVD films were discussed fromthe perspective of sub-monolayers. As a conclusion, we found that it is important to suppress the formation ofsub-monolayers on the substrate to completely form the first layer.
Discharge plasma formed in aqueous solutions has attracted much attention for its applications in environmental purification and material syntheses. The onset and evolution of the discharge plasma in an aqueous solution and transient reactive species formed in it are successfully monitored with micrometer spatial resolution and nanosecond temporal resolution. The combination of a custom-made microscopic discharge system and a high-speed camera provides direct evidence that water vapor bubbles form before the discharge with the thermal phase transition of aqueous solution at the electrode tip. The water vapor bubbles, i.e., locally formed space in the gas phase, connect the gap between the tips of the opposed electrodes. The local gas area formed in aqueous solution plays a crucial role in the ignition and continuance of the discharge plasma. It is also found that the initially formed plasma lasts for under 100 ns and quenches rapidly. However, plasma regenerates in the water vapor bubble and successively bridges the opposing electrodes during the pulsed-voltage application (ca. 1 μs). These two temporally distinct generations of plasma, i.e., the initial plasma (IP) and the following successive plasma (SP), can be seen to correspond to the dielectric breakdown and glow-like plasma, respectively. These results provide an important picture for the proposed mechanism for plasma evolution in water and also important information for the efficient control of the discharge plasma with its applications in waste-water treatments, nanomaterial syntheses with plasma oxidation–reduction reactions, and the chemical modification of the material surfaces in aqueous solutions as a form of “green chemistry.”
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