In utero electroporation and cranial window implantation for in vivo wide-field two-photon calcium imaging using G-CaMP9a transgenic mice

Masayuki Sakamoto; Keisuke Ota; Yayoi Kondo; Michiko Okamura; Hajime Fujii; Haruhiko Bito

doi:10.1016/j.xpro.2022.101421

In utero electroporation and cranial window implantation for in vivo wide-field two-photon calcium imaging using G-CaMP9a transgenic mice

STAR Protoc. 2022 Jun 7;3(2):101421. doi: 10.1016/j.xpro.2022.101421. eCollection 2022 Jun 17.

Authors

Masayuki Sakamoto^{1

2

3}, Keisuke Ota¹, Yayoi Kondo¹, Michiko Okamura¹, Hajime Fujii¹, Haruhiko Bito^{1

4}

Affiliations

¹ Department of Neurochemistry, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
² Department of Optical Neural and Molecular Physiology, Graduate School of Biostudies, Kyoto University, Kyoto, Kyoto 606-8507, Japan.
³ Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency, Kyoto 606-8507, Japan.
⁴ International Research Center for Neurointelligence (WPI-IRCN), The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.

Abstract

We present a protocol to prepare mouse cranial window implantation for in vivo two-photon wide-field calcium imaging. This protocol uses G-CaMP9a transgenic mice, which express a genetically encoded calcium indicator with high signal-to-noise ratio. We describe in utero electroporation, followed by headplate fixation and cranial window implantation. This protocol can be used for measuring neural activity and is suitable for long-term imaging in large populations. Moreover, this protocol does not require preparation of Flp-expressing transgenic mice. For complete details on the use and execution of this protocol, please refer to Sakamoto et al. (2022).

Keywords: Developmental biology; Microscopy; Model Organisms; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Calcium, Dietary
Diagnostic Imaging
Electroporation*
Mice
Mice, Transgenic
Skull* / surgery

Substances

Calcium, Dietary