Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning

Abstract We present a miniature head mounted two-photon fiber-coupled microscope (2P-FCM) for neuronal imaging with active axial focusing enabled using a miniature electrowetting lens. We show three-dimensional two-photon imaging of neuronal structure and record neuronal activity from GCaMP6s fluore...

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Autores principales:	Baris N. Ozbay, Gregory L. Futia, Ming Ma, Victor M. Bright, Juliet T. Gopinath, Ethan G. Hughes, Diego Restrepo, Emily A. Gibson
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2ef9a8fab2784af381b67b48283922d0
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spelling	oai:doaj.org-article:2ef9a8fab2784af381b67b48283922d02021-12-02T15:08:19ZThree dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning10.1038/s41598-018-26326-32045-2322https://doaj.org/article/2ef9a8fab2784af381b67b48283922d02018-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-26326-3https://doaj.org/toc/2045-2322Abstract We present a miniature head mounted two-photon fiber-coupled microscope (2P-FCM) for neuronal imaging with active axial focusing enabled using a miniature electrowetting lens. We show three-dimensional two-photon imaging of neuronal structure and record neuronal activity from GCaMP6s fluorescence from multiple focal planes in a freely-moving mouse. Two-color simultaneous imaging of GFP and tdTomato fluorescence is also demonstrated. Additionally, dynamic control of the axial scanning of the electrowetting lens allows tilting of the focal plane enabling neurons in multiple depths to be imaged in a single plane. Two-photon imaging allows increased penetration depth in tissue yielding a working distance of 450 μm with an additional 180 μm of active axial focusing. The objective NA is 0.45 with a lateral resolution of 1.8 μm, an axial resolution of 10 μm, and a field-of-view of 240 μm diameter. The 2P-FCM has a weight of only ~2.5 g and is capable of repeatable and stable head-attachment. The 2P-FCM with dynamic axial scanning provides a new capability to record from functionally distinct neuronal layers, opening new opportunities in neuroscience research.Baris N. OzbayGregory L. FutiaMing MaVictor M. BrightJuliet T. GopinathEthan G. HughesDiego RestrepoEmily A. GibsonNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-14 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Baris N. Ozbay Gregory L. Futia Ming Ma Victor M. Bright Juliet T. Gopinath Ethan G. Hughes Diego Restrepo Emily A. Gibson Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning
description	Abstract We present a miniature head mounted two-photon fiber-coupled microscope (2P-FCM) for neuronal imaging with active axial focusing enabled using a miniature electrowetting lens. We show three-dimensional two-photon imaging of neuronal structure and record neuronal activity from GCaMP6s fluorescence from multiple focal planes in a freely-moving mouse. Two-color simultaneous imaging of GFP and tdTomato fluorescence is also demonstrated. Additionally, dynamic control of the axial scanning of the electrowetting lens allows tilting of the focal plane enabling neurons in multiple depths to be imaged in a single plane. Two-photon imaging allows increased penetration depth in tissue yielding a working distance of 450 μm with an additional 180 μm of active axial focusing. The objective NA is 0.45 with a lateral resolution of 1.8 μm, an axial resolution of 10 μm, and a field-of-view of 240 μm diameter. The 2P-FCM has a weight of only ~2.5 g and is capable of repeatable and stable head-attachment. The 2P-FCM with dynamic axial scanning provides a new capability to record from functionally distinct neuronal layers, opening new opportunities in neuroscience research.
format	article
author	Baris N. Ozbay Gregory L. Futia Ming Ma Victor M. Bright Juliet T. Gopinath Ethan G. Hughes Diego Restrepo Emily A. Gibson
author_facet	Baris N. Ozbay Gregory L. Futia Ming Ma Victor M. Bright Juliet T. Gopinath Ethan G. Hughes Diego Restrepo Emily A. Gibson
author_sort	Baris N. Ozbay
title	Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning
title_short	Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning
title_full	Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning
title_fullStr	Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning
title_full_unstemmed	Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning
title_sort	three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/2ef9a8fab2784af381b67b48283922d0
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Three dimensional two-photon brain imaging in freely moving mice using a miniature fiber coupled microscope with active axial-scanning

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