In vivo two-photon imaging of mouse hippocampal neurons in dentate gyrus using a light source based on a high-peak power gain-switched laser diode

Biomedical Optics Express
Ryosuke KawakamiTomomi Nemoto

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-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.

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Citations

Feb 26, 2016·Frontiers in Immunology·Enrica PietronigroGabriela Constantin
Apr 21, 2018·Biomedical Optics Express·Keisuke TodaKatsumi Midorikawa
Mar 28, 2017·F1000Research·James N SleighGiampietro Schiavo
Jul 19, 2017·Biomedical Optics Express·Matthias EiblRobert Huber
Oct 20, 2018·Biomedical Optics Express·Gregorio L GaliñanesDaniel Huber
Aug 15, 2018·Journal of Integrative Neuroscience·Satoshi Iwasaki, Yuji Ikegaya
May 9, 2019·Biomedical Optics Express·Matthias EiblSebastian Karpf
Jun 17, 2020·Applied Optics·Yusaku HaraFumihiko Kannari
May 16, 2019·Nature Methods·Feifei WangHongjie Dai
Oct 21, 2020·Journal of Anatomy·Chayanit ChaweewannakornMasahiro Tsuchiya
Dec 20, 2020·Journal of Neuroscience Methods·Yajie Liang, Piotr Walczak
Dec 29, 2020·IScience·Kouichirou IijimaTomomi Nemoto
Jan 18, 2018·Current Opinion in Biomedical Engineering·David R MillerAndrew K Dunn
Sep 3, 2020·ACS Central Science·Shangfeng WangFan Zhang

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