蓝色转移的基因编码的Ca2+指标具有增强的两光子吸收
Abhi Aggarwal1,2,3,4, Smrithi Sunil2, Imane Bendifallah5
1University of Alberta, Department of Chemistry, Edmonton, Alberta, Canada.
Neurophotonics
|April 5, 2024
概括
我们开发了T-GECO1,一种新的蓝色转移的基因编码指标 (GECI). T-GECO1提供了增强的两光子亮度,以改善神经元成像,并在检测动能方面超过现有的GECI.
科学领域:
- 神经科学是一个神经科学.
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 基因编码离子 (GECI) 指标对于监测生物系统中的细胞内动态至关重要.
- 对于神经元活动成像的现有GECI是有限的,需要具有独特光物理性质的新变体.
- 蓝色转移的GECI对于增强的双光子亮度特别有希望,促进了多光子显微镜.
研究的目的:
- 开发和描述T-GECO1,一个高性能蓝移GECI.
- 扩大GECI的调色板,用于神经元活动的先进光成像.
- 评估T-GECO1在各种实验环境中的实用性,包括体外,体内和体外应用.
主要方法:
- 蛋白质工程和定向进化被用来开发T-GECO1.1.
- 纯化的T-GECO1蛋白因其光物理性质而具有特征.
- 在培养的老鼠海马神经元,小鼠大脑和使用一光子和两光子显微镜的ex vivo海马切片中评估了性能.
主要成果:
- T-GECO1在468nm处呈现激发峰值,在500nm处呈现辐射峰值,具有高量子产率 (0.83).
- 它的两光子亮度大约是EGFP的两倍,检测的动态范围是EGFP的15倍.
- T-GECO1在动作潜能检测方面表现出优异的信号噪声比,相比于在两光子激发下的后一代GCaMP变体.
结论:
- T-GECO1是一款高性能蓝转移的GECI.
- 它比现有的GCaMP变体在神经元活动成像方面具有显著的优势,特别是在两光子激发下.
- T-GECO1扩展了先进的体外和体内神经科学研究工具包.
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