Three-photon in vivo imaging of neurons and glia in the medial prefrontal cortex with sub-cellular resolution
The medial prefrontal cortex (mPFC) is important for higher cognitive functions, including working memory, decision making, and emotional control. In vivo recordings of neuronal activity in the mPFC have been achieved via invasive electrical and optical approaches. Here we apply low invasive three-photon in vivo imaging in the mPFC of the mouse at unprecedented depth. Specifically, we measure neuronal and astrocytic Ca<sup>2+</sup>-transient parameters in awake head-fixed mice up to a depth of 1700 µm. Furthermore, we longitudinally record dendritic spine density (0.41 ± 0.07 µm<sup>-1</sup>) deeper than 1 mm for a week. Using 1650 nm wavelength to excite red fluorescent microglia, we quantify their processes' motility (58.9 ± 2% turnover rate) at previously unreachable depths (1100 µm). We establish three-photon imaging of the mPFC enabling neuronal and glial recordings with subcellular resolution that will pave the way for novel discoveries in this brain region.
Authors
- Ivy Chi Wai Chan (ORCID: https://orcid.org/0000-0002-0261-1900)
- Eleonora Ambrad Giovannetti (ORCID: https://orcid.org/0000-0003-0420-154X)
- Nicole Reichenbach
- Gabor C. Petzold (ORCID: https://orcid.org/0000-0002-0145-8641)
- Stefanie Poll (ORCID: https://orcid.org/0000-0001-5301-2791)
- Barbara Schaffran
- Manuel Mittag (ORCID: https://orcid.org/0000-0001-9908-3464)
- Hans‐Ulrich Fried (ORCID: https://orcid.org/0000-0001-7557-1199)
- Stefan Lindén (ORCID: https://orcid.org/0000-0002-1028-9528)
- Sanjeev Kumar Kaushalya (ORCID: https://orcid.org/0000-0002-3469-2637)
- Falko Fuhrmann
- Alex Simon Lagurin
- Mónika Müller
- Fabrizio Musacchio (ORCID: https://orcid.org/0000-0002-9043-3349)
- Gaia Tavosanis (ORCID: https://orcid.org/0000-0002-8679-5515)
- Emily R. Burnside (ORCID: https://orcid.org/0000-0003-3238-0602)
- Martin Fuhrmann (ORCID: https://orcid.org/0000-0001-7672-2913)
- Felix Christopher Nebeling (ORCID: https://orcid.org/0000-0002-2749-7163)
- Frank Bradke (ORCID: https://orcid.org/0000-0002-0345-3772)
- Michael Maibach (ORCID: https://orcid.org/0009-0002-9949-5463)
Institutions
- University of Bonn (DE)
- University Hospital Bonn (DE)
- German Center for Neurodegenerative Diseases (DE)
- NeoPhotonics (United States) (US)
- RWTH Aachen University (DE)
Publication Details
- Journal
- Communications Biology
- Published
- 2025-05-23
- DOI
- https://doi.org/10.1038/s42003-025-08079-8
- Citations
- 5
- Primary Topic
- Neuroinflammation and Neurodegeneration Mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 4.21