Closed-loop two-photon functional imaging in a freely moving animal

Paul McNulty, Rui Wu, Akihiro Yamaguchi, Ellie S. Heckscher, Andrew Haas, Amajindi Nwankpa, Mirna Mihovilovic Skanata () and and Marc Gershow ()
Additional contact information
Paul McNulty: New York University
Rui Wu: New York University
Akihiro Yamaguchi: New York University
Ellie S. Heckscher: University of Chicago
Andrew Haas: New York University
Amajindi Nwankpa: New York University
Mirna Mihovilovic Skanata: New York University
and Marc Gershow: New York University

Nature Communications, 2025, vol. 16, issue 1, 1-22

Abstract: Abstract Direct measurement of neural activity in freely moving animals is essential for understanding how the brain controls and represents behaviors. Genetically encoded calcium indicators report neural activity as changes in fluorescence intensity, but brain motion confounds quantitative measurement of fluorescence. Translation, rotation, and deformation of the brain and the movements of intervening scattering or autofluorescent tissue all alter the amount of fluorescent light captured by a microscope. Compared to single-photon approaches, two-photon microscopy is less sensitive to scattering and off-target fluorescence, but more sensitive to motion, and two photon imaging has always required anchoring the microscope to the brain. We developed a closed-loop resonant axial-scanning high-speed two-photon (CRASH2p) microscope for real-time 3D motion correction in unrestrained animals, without implantation of reference markers. We complemented CRASH2p with a ‘Pong’ scanning strategy and a multi-stage registration pipeline. We performed volumetric ratiometrically corrected functional imaging in the CNS of freely moving Drosophila larvae and discovered previously unknown neural correlates of behavior.

Date: 2025
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-025-60648-x Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-60648-x

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-025-60648-x

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().