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一小时,千赫兹采样速率 3D 单个病毒在活细胞中的跟踪通过StayGold光蛋白融合实现

Yuxin Lin¹, Jack Exell¹, Haoting Lin¹

¹Department of Chemistry, Duke University; Durham, North Carolina 27708, United States.

bioRxiv : the preprint server for biology

|April 1, 2024

概括

新的StayGold光蛋白可以在活细胞中进行长达1小时的高速3D单个病毒追踪 (SVT). 这一突破允许详细研究病毒动态和整个病毒生命周期.

科学领域:

生物物理学的生物物理.
细胞生物学细胞生物学
病毒学病毒学

背景情况:

病毒感染涉及复杂的时空动态.
长期,高分辨率的病毒跟踪需要明亮,光稳定的光标签.
现有的光蛋白限制了病毒追踪研究的持续时间和分辨率.

研究的目的:

为高性能单个病毒跟踪 (SVT) 开发先进的标签策略.
为了利用光稳定的StayGold光蛋白进行扩展的病毒动态研究.
为了使 kHz 采样率能够在活细胞中进行长达一小时的 3D SVT.

主要方法:

产生与StayGold光蛋白 (StayGold-VLPs) 融合的病毒样颗粒.
在活细胞中实施3D单个病毒追踪 (SVT).
实现kHz采样速率用于长时间的病毒轨迹分析.

主要成果:

与现有变种相比,StayGold-VLPs表现出优越的光稳定性和信号强度.
启用了一个小时的3D SVT与kHz采样率,显著扩大跟踪时间.
从单个病毒轨迹收集了指数级更多的数据.

结论:

StayGold-VLP提供了一个简单的,可通用的标签策略,以提高SVT的性能.
扩大的光子预算有助于对病毒贩运动态进行全面研究.
这种方法为观察单个病毒的完整生命周期铺平了道路.

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