OSCR

Simultaneous two- and three-photon multiplane imaging across cortical layers in freely moving mice.

Overview

Authors: Alexandr Klioutchnikov1, Damian J Wallace1, Caleb Berdahl1, Adam Sugi1, Juergen Sawinski1, Jason N D Kerr1
ORCID iDs: Alexandr Klioutchnikov, Damian J Wallace, Caleb Berdahl, Juergen Sawinski, Jason N D Kerr
  1. Department of Behavior and Brain Organization, Max Planck Institute for Neurobiology of Behavior - caesar, Bonn, Germany
Institutions: Max Planck Institute for Neurobiology of Behavior – caesar (Germany)
Journal: Nature methods, volume 23, issue 7, pages 1437-1446
Dates: received 6 May 2025; accepted 6 May 2026; published online 9 June 2026; in print 2026
Type: Research article · Language: English
License: CC BY
Identifiers: DOI 10.1038/s41592-026-03125-7 · PMID 42265209 · PMCID PMC13345954 · OpenAlex W7164016784
Open access: hybrid, a free copy (OpenAlex)
Status: code on request
Categories: histology / microscopy (modality), optical imaging (calcium, voltage, 2-photon) (modality), mouse (organism), systems (subfield)
Methods: Spectral & time-frequency, Connectivity, Statistics, Preprocessing, Graphs, fMRI & imaging, Single-unit activity, calcium imaging, Smoothing, state filtering, decompositions
Keywords: Multiphoton microscopy, Visual system
MeSH: Microscopy, Fluorescence, Multiphoton*, Neurons*, Parietal Lobe*, Visual Cortex*, Animals, Male, Mice, Mice, Inbred C57BL (* major topic)
Topic: Advanced Fluorescence Microscopy Techniques (Biophysics, Biochemistry, Genetics and Molecular Biology), according to OpenAlex
Citations: not cited yet (Europe PMC); 72 references in the paper

Abstract

Head-mounted multiphoton microscopes enable imaging of activity from neuronal populations spread throughout the cortical layers in freely moving mice but so far have been restricted to recording from one cortical layer at a time. Here, combining two- and three-photon-based excitation delivered through multiple fibers, we built a head-mounted multiplane microscope enabling near-simultaneous imaging of neuronal activity from five vertically separated planes, spread across multiple cortical layers. Both excitation pathways had remote focusing mechanisms for fine axial adjustments, enabling activity recordings from the same neuronal populations over weeks in freely behaving mice. The lightweight microscope utilized an onboard, two-channel detection system designed to enable activity recordings from neuronal populations spread across visual-cortex layers in both lit and dark conditions as well as imaging activity across posterior parietal cortex layers during complex gap-crossing behaviors. We show that during gap-crossing tasks, layer 5 and 2/3 neuronal subpopulations in the posterior parietal cortex have differential pattern sequences during free decision-making.

Reproduced under the paper's license (CC BY), from the paper cited above.

Code

The paper says that its authors' code is available on request: it was not published with the paper, so there is nothing to verify.

Code availability

The code used to pre-process data, including de-mixing of imaging channels is available in the following Edmond (Max Planck Society) repository (10.17617/3.NW9DW0).

Reproduced under the paper's license (CC BY), from the paper cited above.

Tracing map

A tracing map links a paper to the code its authors published: this paper has none (its code is available on request), so it has no map.

Data

No dataset and no data link were found in the paper.

Data availability

The imaging data split into individual planes as well as tracked behavioral data are available in an Edmond (Max Planck Society) repository (10.17617/3.NW9DW0). The full raw data will be available on request due to large volumes.

Reproduced under the paper's license (CC BY), from the paper cited above.

Versions

The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.

Version 2, 28 September 2026

  • Publisher: n/a → Nature Portfolio

Version 1, 27 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 6 authors, 2 keywords, 8 MeSH terms, 68 references.

Cite

This paper

Klioutchnikov, A., Wallace, D. J., Berdahl, C., Sugi, A., Sawinski, J., & Kerr, J. N. D. (2026). Simultaneous two- and three-photon multiplane imaging across cortical layers in freely moving mice. Nature methods, 23(7), 1437-1446. https://doi.org/10.1038/s41592-026-03125-7

BibTeX

@article{klioutchnikov2026simultaneous,
author = {Klioutchnikov, Alexandr and Wallace, Damian J and Berdahl, Caleb and Sugi, Adam and Sawinski, Juergen and Kerr, Jason N D},
title = {{Simultaneous two- and three-photon multiplane imaging across cortical layers in freely moving mice}},
journal = {Nature methods},
year = {2026},
month = jun,
volume = {23},
number = {7},
pages = {1437--1446},
publisher = {Nature Portfolio},
issn = {1548-7091},
doi = {10.1038/s41592-026-03125-7},
url = {https://doi.org/10.1038/s41592-026-03125-7},
pmid = {42265209},
pmcid = {PMC13345954}
}

RIS

TY - JOUR
AU - Klioutchnikov, Alexandr
AU - Wallace, Damian J
AU - Berdahl, Caleb
AU - Sugi, Adam
AU - Sawinski, Juergen
AU - Kerr, Jason N D
TI - Simultaneous two- and three-photon multiplane imaging across cortical layers in freely moving mice
T2 - Nature methods
J2 - Nat Methods
PY - 2026
DA - 2026/06/09
VL - 23
IS - 7
SP - 1437
EP - 1446
SN - 1548-7091
PB - Nature Portfolio
DO - 10.1038/s41592-026-03125-7
UR - https://doi.org/10.1038/s41592-026-03125-7
LA - en
ER -

CSL-JSON

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