In utero electroporation and cranial window implantation for in vivo wide-field two-photon calcium imaging using G-CaMP9a transgenic mice.
In utero electroporation and cranial window implantation for in vivo wide-field two-photon calcium imaging using G-CaMP9a transgenic mice.
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子宫内电穿孔和颅窗植入用于使用G-CaMP 9a转基因小鼠的体内宽场双光子钙成像。
DOI：
10.1016/j.xpro.2022.101421
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发表时间：
2022-06-17
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我们提出了一个协议，准备在体内双光子宽场钙成像小鼠颅窗植入。该方案使用G-CaMP 9a转基因小鼠，其表达具有高信噪比的遗传编码的钙指示剂。我们描述了子宫内电穿孔，然后进行头板固定和颅窗植入。该方案可用于测量神经活动，适用于大规模人群的长期成像。此外，该方案不需要制备表达Flp的转基因小鼠。有关本方案使用和执行的完整详细信息，请参阅。用于体内钙成像的G-CaMP 9a基因敲入小鼠的制备在子宫内电穿孔以将Flp重组酶引入G-CaMP 9a小鼠中用于体内双光子钙成像的颅窗植入出版商说明：进行任何实验方案都需要遵守当地机构关于实验室安全性和伦理学的指南。我们提出了一个协议，准备在体内双光子宽场钙成像小鼠颅窗植入。该方案使用G-CaMP 9a转基因小鼠，其表达具有高信噪比的遗传编码的钙指示剂。我们描述了子宫内电穿孔，然后进行头板固定和颅窗植入。该方案可用于测量神经活动，适用于大规模人群的长期成像。此外，该方案不需要制备表达Flp的转基因小鼠。
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. Preparation of G-CaMP9a knock-in mice for in vivo calcium imaging In utero electroporation to introduce Flp recombinase in G-CaMP9a mice Implantation of cranial window for in vivo two-photon calcium imaging Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.