使用用于双光子显微镜的快速指示器进行持续深层组织电压记录
Zhuohe Liu1, Xiaoyu Lu2, Vincent Villette3
1Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005, USA.
Cell
|August 19, 2022
概括
我们开发了JEDI-2P, 一种新的遗传编码电压指示器, 显著提高了双光子显微镜的性能,
科学领域:
- 神经科学
- 生物技术
- 显微镜
背景情况:
- 基因编码的电压指示器 (GEVIs) 对于特定电池类型的电压动态监测至关重要.
- 目前的GEVI表现出性能上的局限性,特别是在双光子显微镜下,阻碍了深层组织的神经记录.
研究的目的:
- 用两光子显微镜优化GEVI以提高性能.
- 开发一个新的GEVI,JEDI-2P,提高速度,亮度,灵敏度和光稳定性.
主要方法:
- 开发一个多参数高吞吐量平台来优化GEVI.
- 应用该平台来识别和描述JEDI-2P.
- 在Drosophila,小鼠视网膜和清醒的小鼠中对JEDI-2P的体内和体外验证.
主要成果:
- 与现有的指标相比,JEDI-2P显示出更高的速度,亮度,灵敏度和光稳定性.
- 在Drosophila内部神经元和小鼠视网膜皮细胞中成功报告了光唤起的反应.
- 通过共振扫描和ULoVE显微镜在清醒的小鼠中实现单个皮质神经元的长期光学记录.
- 用JEDI-2P进行的ULoVE记录检测到深度超过400μm的尖峰,并揭示了神经元电压的相关性.
结论:
- JEDI-2P代表了GEVI在双光子显微镜中的重大进步.
- 这种指标可以在深度神经回路中进行高分辨率电压成像.
- 在体内研究神经电路功能的新途径.
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