Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection

We recently established a simple and versatile adeno-associated virus (AAV) induction approach that enables dense (>90% labeled neurons) and cortical-wide Ca(2+) sensor expression. Here, we describe the stepwise protocol for neonatal AAV injection of a Ca(2+) sensor. We also detail the steps for...

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Autores principales: Oomoto, Ikumi, Uwamori, Hiroyuki, Matsubara, Chie, Odagawa, Maya, Kobayashi, Midori, Kobayashi, Kenta, Ota, Keisuke, Murayama, Masanori
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672102/
https://www.ncbi.nlm.nih.gov/pubmed/34950887
http://dx.doi.org/10.1016/j.xpro.2021.101007
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author Oomoto, Ikumi
Uwamori, Hiroyuki
Matsubara, Chie
Odagawa, Maya
Kobayashi, Midori
Kobayashi, Kenta
Ota, Keisuke
Murayama, Masanori
author_facet Oomoto, Ikumi
Uwamori, Hiroyuki
Matsubara, Chie
Odagawa, Maya
Kobayashi, Midori
Kobayashi, Kenta
Ota, Keisuke
Murayama, Masanori
author_sort Oomoto, Ikumi
collection PubMed
description We recently established a simple and versatile adeno-associated virus (AAV) induction approach that enables dense (>90% labeled neurons) and cortical-wide Ca(2+) sensor expression. Here, we describe the stepwise protocol for neonatal AAV injection of a Ca(2+) sensor. We also detail the steps for subsequent craniotomy to generate a chronic cranial window, followed by wide-field two-photon Ca(2+) imaging in an awake mouse. This protocol serves as an alternative to the use of transgenic animals and offers translatable options for cortical-wide experiments. For complete details on the use and execution of this protocol, please refer to Ota et al. (2021).
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spelling pubmed-86721022021-12-22 Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection Oomoto, Ikumi Uwamori, Hiroyuki Matsubara, Chie Odagawa, Maya Kobayashi, Midori Kobayashi, Kenta Ota, Keisuke Murayama, Masanori STAR Protoc Protocol We recently established a simple and versatile adeno-associated virus (AAV) induction approach that enables dense (>90% labeled neurons) and cortical-wide Ca(2+) sensor expression. Here, we describe the stepwise protocol for neonatal AAV injection of a Ca(2+) sensor. We also detail the steps for subsequent craniotomy to generate a chronic cranial window, followed by wide-field two-photon Ca(2+) imaging in an awake mouse. This protocol serves as an alternative to the use of transgenic animals and offers translatable options for cortical-wide experiments. For complete details on the use and execution of this protocol, please refer to Ota et al. (2021). Elsevier 2021-12-10 /pmc/articles/PMC8672102/ /pubmed/34950887 http://dx.doi.org/10.1016/j.xpro.2021.101007 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Oomoto, Ikumi
Uwamori, Hiroyuki
Matsubara, Chie
Odagawa, Maya
Kobayashi, Midori
Kobayashi, Kenta
Ota, Keisuke
Murayama, Masanori
Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection
title Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection
title_full Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection
title_fullStr Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection
title_full_unstemmed Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection
title_short Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection
title_sort protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672102/
https://www.ncbi.nlm.nih.gov/pubmed/34950887
http://dx.doi.org/10.1016/j.xpro.2021.101007
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