Quicke, P;
Howe, CL;
Song, P;
Jadan, HV;
Song, C;
Knöpfel, T;
Neil, M;
... Foust, AJ; + view all
(2020)
Subcellular resolution three-dimensional light-field imaging with genetically encoded voltage indicators.
Neurophotonics
, 7
(3)
, Article 035006. 10.1117/1.NPh.7.3.035006.
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Abstract
Significance: Light-field microscopy (LFM) enables high signal-to-noise ratio (SNR) and light efficient volume imaging at fast frame rates. Voltage imaging with genetically encoded voltage indicators (GEVIs) stands to particularly benefit from LFM's volumetric imaging capability due to high required sampling rates and limited probe brightness and functional sensitivity. Aim: We demonstrate subcellular resolution GEVI light-field imaging in acute mouse brain slices resolving dendritic voltage signals in three spatial dimensions. Approach: We imaged action potential-induced fluorescence transients in mouse brain slices sparsely expressing the GEVI VSFP-Butterfly 1.2 in wide-field microscopy (WFM) and LFM modes. We compared functional signal SNR and localization between different LFM reconstruction approaches and between LFM and WFM. Results: LFM enabled three-dimensional (3-D) localization of action potential-induced fluorescence transients in neuronal somata and dendrites. Nonregularized deconvolution decreased SNR with increased iteration number compared to synthetic refocusing but increased axial and lateral signal localization. SNR was unaffected for LFM compared to WFM. Conclusions: LFM enables 3-D localization of fluorescence transients, therefore eliminating the need for structures to lie in a single focal plane. These results demonstrate LFM's potential for studying dendritic integration and action potential propagation in three spatial dimensions.
| Type: | Article |
|---|---|
| Title: | Subcellular resolution three-dimensional light-field imaging with genetically encoded voltage indicators |
| Location: | United States |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1117/1.NPh.7.3.035006 |
| Publisher version: | https://doi.org/10.1117/1.NPh.7.3.035006 |
| Language: | English |
| Additional information: | This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
| Keywords: | genetically encoded voltage indicator, light-field microscopy, voltage imaging |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Electronic and Electrical Eng |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10110571 |
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