Two-photon calcium imaging of the medial prefrontal cortex and hippocampus without cortical invasion
Abstract
In vivo two-photon calcium imaging currently allows us to observe the activity of multiple neurons up to ~900 µm below the cortical surface without cortical invasion. However, many important brain areas are located deeper than this. Here, we used an 1100 nm laser that underfilled the back aperture of the objective together with red genetically encoded calcium indicators to establish two-photon calcium imaging of the intact mouse brain and detect neural activity up to 1200 μm from the cortical surface. This imaging was obtained from the medial prefrontal cortex (the prelimbic area) and the hippocampal CA1 region. We found that neural activity before water delivery repeated at a constant interval was higher in the prelimbic area than in layer 2/3 of the secondary motor area. Reducing the invasiveness of imaging is an important strategy to reveal the intact brain processes active in cognition and memory.
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Funding
Ministry of Education, Culture, Sports, Science, and Technology (Grant-in-Aids for Scientific Research on Innovative Areas (15H01455))
- Masanori Matsuzaki
Ministry of Education, Culture, Sports, Science, and Technology (Grant-in-Aids for Scientific Research on Innovative Areas (17H06309))
- Masanori Matsuzaki
Ministry of Education, Culture, Sports, Science, and Technology (Grant-in-Aids for Scientific Research (A) (15H02350))
- Masanori Matsuzaki
Takeda Science Foundation
- Masanori Matsuzaki
Japan Agency for Medical Research and Development (The Strategic Research Program for Brain Sciences)
- Masanori Matsuzaki
Japan Agency for Medical Research and Development (The program for Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS))
- Masanori Matsuzaki
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All animal experiments were approved by the Institutional Animal Care and Use Committee of The University of Tokyo, Japan (Medicine-P16-012).
Copyright
© 2017, Kondo et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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