Subcellular resolution three-dimensional light-field imaging with genetically encoded voltage indicators
Author(s)
Type
Journal Article
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.
Date Issued
2020-08-28
Date Acceptance
2020-08-07
Citation
Neurophotonics, 2020, 7 (3)
ISSN
2329-4248
Publisher
Society of Photo-optical Instrumentation Engineers
Journal / Book Title
Neurophotonics
Volume
7
Issue
3
Copyright Statement
©The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License.Distribution or reproduction of this work in whole or in part requires full attribution of the original pub-lication, including its DOI.[DOI:10.1117/1.NPh.7.3.035006]
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/32904628
PII: 20044R
Subjects
genetically encoded voltage indicator
light-field microscopy
voltage imaging
Publication Status
Published
Coverage Spatial
United States
Article Number
ARTN 035006