Real-time 3D movement correction for two-photon imaging in behaving animals

Author(s):Show less -

Griffiths, Victoria A. ^[1] ; Valera, Antoine M. ^[1] ; Lau, Joanna Y. N. ^[1] ; Ros, Hana ^[1] ; Younts, Thomas J. ^[1] ; Marin, Boris ^{[1, 2]} ; Baragli, Chiara ^[1] ; Coyle, Diccon ^[1] ; Evans, Geoffrey J. ^{[1, 3]} ; Konstantinou, George ^{[4, 1]} ; Koimtzis, Theo ^{[1, 5]} ; Nadella, K. M. Naga Srinivas ^[1] ; Punde, Sameer A. ^[1] ; Kirkby, Paul A. ^[1] ; Bianco, Isaac H. ^[1] ; Silver, R. Angus ^[1]

Affiliation

^[1] UCL, Dept Neurosci Physiol & Pharmacol, London - England

^[2] Univ Fed ABC, Ctr Matemat Comp & Cognicao, Sao Bernardo Do Campo - Brazil

^[3] Sencon UK Ltd, Dept Engn, Droitwich - England

^[4] Francis Crick Inst, London - England

^[5] Opt Metrol Serv, Stansted - England

• Document type: Journal article
• Source: NATURE METHODS; v. 17, n. 7 JUN 2020.
• Web of Science Citations: 0

Abstract

Two-photon microscopy is widely used to investigate brain function across multiple spatial scales. However, measurements of neural activity are compromised by brain movement in behaving animals. Brain motion-induced artifacts are typically corrected using post hoc processing of two-dimensional images, but this approach is slow and does not correct for axial movements. Moreover, the deleterious effects of brain movement on high-speed imaging of small regions of interest and photostimulation cannot be corrected post hoc. To address this problem, we combined random-access three-dimensional (3D) laser scanning using an acousto-optic lens and rapid closed-loop field programmable gate array processing to track 3D brain movement and correct motion artifacts in real time at up to 1 kHz. Our recordings from synapses, dendrites and large neuronal populations in behaving mice and zebrafish demonstrate real-time movement-corrected 3D two-photon imaging with submicrometer precision. Real-time 3D movement correction by tracking a fluorescent bead in the field of view enables functional imaging with 3D two-photon random-access microscopy in behaving mice and zebrafish. (AU)

• FAPESP's process: 18/20277-0 - Computational and systems neuroscience
• Grantee: Antonio Carlos Roque da Silva Filho
• Support Opportunities: Regular Research Grants