Quasi-simultaneous multiplane calcium imaging of neuronal circuits

Chong, E. Z., Panniello, M., Barreiros, I., Kohl, M. M. and Booth, M. J. (2019) Quasi-simultaneous multiplane calcium imaging of neuronal circuits. Biomedical Optics Express, 10(1), pp. 267-282. (doi: 10.1364/BOE.10.000267) (PMID:30775099) (PMCID:PMC6363184)

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Two-photon excitation fluorescence microscopy is widely used to study the activity of neuronal circuits. However, the fast imaging is typically constrained to a single lateral plane for a standard microscope design. Given that cortical neuronal networks in a mouse brain are complex three-dimensional structures organised in six histologically defined layers which extend over many hundreds of micrometres, there is a strong demand for microscope systems that can record neuronal signalling in volumes. Henceforth, we developed a quasi-simultaneous multiplane imaging technique combining acousto-optic deflector and static remote focusing to provide fast imaging of neurons from different axial positions inside the cortical layers without the need for the mechanical interference of either the objective lens or the specimen. The hardware and the software are easily adaptable to existing two-photon microscopes. Here, we demonstrated that our imaging method can record, at high speed and high image contrast, the calcium dynamics of neurons in two different imaging planes separated axially, with the in-focus and the refocused planes 120 μm and 250 μm below the brain surface respectively.

Item Type:Articles
Additional Information:This work was supported by a Wellcome Trust Seed Award (109908/Z/15/Z) to MMK and a Human Frontiers Science Programme Young Investigator Award (RGY0073/2015) to MMK.
Glasgow Author(s) Enlighten ID:Kohl, Dr Michael and Panniello, Dr Mariangela
Authors: Chong, E. Z., Panniello, M., Barreiros, I., Kohl, M. M., and Booth, M. J.
College/School:College of Medical Veterinary and Life Sciences > School of Psychology & Neuroscience
Journal Name:Biomedical Optics Express
Publisher:Optical Society of America
ISSN (Online):2156-7085
Published Online:19 December 2018
Copyright Holders:Copyright © 2019 The Authors
First Published:First published in Biomedical Optics Express 10(1):267-282
Publisher Policy:Reproduced under a Creative Commons License

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