Real-time 3D movement correction for two-photon imaging in behaving animals
| Autor: | Paul Anthony Kirkby, Theo Koimtzis, Antoine M. Valera, R. Angus Silver, George Konstantinou, Geoffrey J. Evans, Chiara Baragli, Hana Roš, Sameer A Punde, Isaac H. Bianco, Joanna Yn Lau, Boris Marin, K. M. Naga Srinivas Nadella, Diccon Coyle, Thomas J. Younts, Victoria A. Griffiths |
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| Rok vydání: | 2020 |
| Předmět: |
Male
Laser scanning Computer science Movement Field of view Tracking (particle physics) Biochemistry Article Photostimulation law.invention Mice 03 medical and health sciences Imaging Three-Dimensional Two-photon excitation microscopy law Microscopy Animals Molecular Biology Zebrafish 030304 developmental biology 0303 health sciences Cell Biology Mice Inbred C57BL Functional imaging Lens (optics) Microscopy Fluorescence Multiphoton Female Neuroscience Biotechnology |
| Zdroj: | Nature methods |
| ISSN: | 1548-7105 1548-7091 |
| Popis: | 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 artefacts are typically corrected using post-hoc processing of 2D 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 3D laser scanning using an acousto-optic lens and rapid closed-loop FPGA 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 sub-micrometer precision. Editor summary: 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. |
| Databáze: | OpenAIRE |
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