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

Ryosuke Kawakami, Kazuaki Sawada, Yuta Kusama, Yi Cheng Fang, Shinya Kanazawa, Yuichi Kozawa, Shunichi Sato, Hiroyuki Yokoyama, Tomomi Nemoto

Research output: Contribution to journalArticle

46 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)891-901
Number of pages11
JournalBiomedical Optics Express
Volume6
Issue number3
DOIs
Publication statusPublished - 2015 Jan 1

ASJC Scopus subject areas

  • Biotechnology
  • Atomic and Molecular Physics, and Optics

Fingerprint Dive into the research topics of 'In vivo two-photon imaging of mouse hippocampal neurons in dentate gyrus using a light source based on a high-peak power gainswitched laser diode'. Together they form a unique fingerprint.

  • Cite this