Journal
BIOMEDICAL OPTICS EXPRESS
Volume 6, Issue 3, Pages 891-901Publisher
OPTICAL SOC AMER
DOI: 10.1364/BOE.6.000891
Keywords
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Funding
- JSPS KAKENHI grant of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan [22113005, 26242082]
- NanoMacro Materials, Devices and System Research Alliance (MEXT)
- Network Joint Research Center for Materials and Devices (MEXT)
- Grants-in-Aid for Scientific Research [22113005, 26242082, 14J01625, 26650107] Funding Source: KAKEN
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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-to-background 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. (C) 2015 Optical Society of America
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