温和的罗达胺用于活细胞光显微镜
Tianyan Liu1,2, Julian Kompa3, Jing Ling1,2
1College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing 100871, China.
ACS central science
|October 28, 2024
概括
新的温和罗达胺可显著降低长期生物成像的光毒性. 这些循环八三烯 (COT) 结合光体使得先进的时间延迟应用能够减少细胞光损伤.
科学领域:
- 生物化学 生物化学
- 细胞生物学 细胞生物学
- 显微镜的使用方法
背景情况:
- 罗达胺染料对生物成像至关重要,但在长时间照明下会产生光毒性,限制了时间延迟研究.
- 光毒性,特别是单片氧生成,是长期活细胞成像应用的重大挑战.
- 现有的罗达胺优化在很大程度上忽视了光毒性问题.
研究的目的:
- 开发具有降低光毒性的新型罗达胺光体,用于增强时差生物成像.
- 引入一种创建"温柔罗达胺"的总体策略,该策略可以保持出色的成像性能,同时最大限度地减少光损伤.
- 为各种细胞成像目标和蛋白质标签创建一个多功能工具包的温和罗达胺.
主要方法:
- 循环环烯 (COT) 与罗达胺支架的结合,以创建新的光体.
- 评估光谱特性,细胞透性和光毒性,包括单片氧形成.
- 在线粒体,DNA,actin和HaloTag标记蛋白质的时间间隔成像中应用COT结合的罗达胺.
主要成果:
- 开发了跨越可见光谱的温和罗达胺,显著降低了光毒性和单片氧形成.
- 与传统的罗达胺相比,证明了保留的光谱特性和细胞透性.
- 展示了各种细胞结构和蛋白质标记的时差成像中的减少细胞光损伤,使得像电压成像这样的应用成为可能.
结论:
- 温和的罗达胺在长期生物成像中为光毒性提供了可行的解决方案.
- 开发的策略为先进的时间缩短显微镜提供了一套多功能工具,可减少光损伤.
- 这些新的光体扩大了活细胞成像的可能性,包括动态过程和电压传感.
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