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Two-photon 3D imaging of optically stimulated neural activity at 100 Hz.

Overview

Authors: Dongli Xu1, Fuu-Jiun Hwang2, Jun B Ding2,3,4, Leilei Peng1
ORCID iDs: Fuu-Jiun Hwang, Jun B Ding, Leilei Peng
  1. Wyant College of Optical Sciences, University of Arizona, Tucson, AZ 85721 USA
  2. Department of Neurosurgery, Stanford University, Stanford, CA 94305 USA
  3. Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA 94305 USA
  4. Wu-Tsai Neuroscience Institute, Stanford University, Stanford, CA 94305 USA
Institutions: University of Arizona (United States); Stanford University (United States); Stanford Medicine (United States)
Journal: Light, science & applications, volume 15, issue 1, article 303
Dates: received 11 December 2025; accepted 11 June 2026; published online 3 July 2026
Type: Research article · Language: English
License: CC BY
Identifiers: DOI 10.1038/s41377-026-02395-2 · PMID 42393079 · PMCID PMC13328388 · OpenAlex W7167047868
Open access: gold, a free copy (OpenAlex)
Status: data only
Categories: optical imaging (calcium, voltage, 2-photon) (modality)
Methods: Preprocessing, Evoked potentials, fMRI & imaging, Single-unit activity, calcium imaging
Keywords: Imaging and sensing, Biophotonics
Topic: Advanced Fluorescence Microscopy Techniques (Biophysics, Biochemistry, Genetics and Molecular Biology), according to OpenAlex
Funding: U.S. Department of Health & Human Services | NIH | National Institute of Biomedical Imaging and Bioengineering (NIBIB) (R56EB034921, R01EB034921); NIBIB NIH HHS (R56 EB034921, R01 EB034921); U.S. Department of Health & Human Services | NIH | National Institute of Biomedical Imaging and Bioengineering (R01EB034921, R56EB034921); NINDS NIH HHS (R01 NS091144)
Citations: not cited yet (Europe PMC); 29 references in the paper

Abstract

Understanding how neurons integrate synaptic inputs requires imaging techniques capable of capturing rapid, three-dimensional dendritic events. These processes occur on millisecond timescales and submicron spatial scales, exceeding the speed of conventional two-photon microscopy (2PM). We developed dual-view Bessel two-photon projection microscopy (dv-B2PM), a high-speed volumetric imaging approach that achieves 100 Hz whole-volume acquisition with synaptic-level resolution. dv-B2PM simultaneously records two orthogonal projections of the same 3D volume, preserving spatial information while minimizing ambiguity from structural overlap. Combining dv-B2PM with two-photon glutamate uncaging, we visualized 3D Ca²⁺ dynamics in neurons following localized stimulation. Multi-timescale analysis revealed dendrite-to-soma Ca²⁺ signal propagation, back propagated Ca²⁺ signal from the soma, and multi-frequency (5–40 Hz) Ca²⁺ transients activated along apical dendrites at speeds from ten of microns per second to millimeters per seconds. These findings demonstrate dv-B2PM as a powerful tool for direct visualization of 3D calcium dynamics associated with dendritic integration across extended neuronal structures, bridging the gap between optical imaging and the dynamic biophysics of neuronal integration.

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

Code

The paper links to its data, not to its authors' code: see the Data section.

The paper's code and data availability statement is in the Data section.

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Data

Datasets cited

Data availability

Image data and scripts used to analyze the data can be downloaded at.https://zenodo.org/records/17887796.

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 1, 27 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 4 authors, 2 keywords, 4 funders, 29 references.

Cite

This paper

Xu, D., Hwang, F.-J., Ding, J. B., & Peng, L. (2026). Two-photon 3D imaging of optically stimulated neural activity at 100 Hz. Light, science & applications, 15(1), 303. https://doi.org/10.1038/s41377-026-02395-2

BibTeX

@article{xu2026two,
author = {Xu, Dongli and Hwang, Fuu-Jiun and Ding, Jun B and Peng, Leilei},
title = {{Two-photon 3D imaging of optically stimulated neural activity at 100 Hz}},
journal = {Light, science \& applications},
year = {2026},
month = jul,
volume = {15},
number = {1},
pages = {303},
publisher = {Nature Publishing Group},
issn = {2095-5545},
doi = {10.1038/s41377-026-02395-2},
url = {https://doi.org/10.1038/s41377-026-02395-2},
pmid = {42393079},
pmcid = {PMC13328388}
}

RIS

TY - JOUR
AU - Xu, Dongli
AU - Hwang, Fuu-Jiun
AU - Ding, Jun B
AU - Peng, Leilei
TI - Two-photon 3D imaging of optically stimulated neural activity at 100 Hz
T2 - Light, science & applications
J2 - Light Sci Appl
PY - 2026
DA - 2026/07/03
VL - 15
IS - 1
SP - 303
SN - 2095-5545
PB - Nature Publishing Group
DO - 10.1038/s41377-026-02395-2
UR - https://doi.org/10.1038/s41377-026-02395-2
LA - en
ER -

CSL-JSON

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