双色相对照光和电子显微镜用于可视化线粒体和溶解体之间的相互作用
1Neural circuit research group, Korea Brain Research Institute; j0312@kbri.re.kr.
Journal of visualized experiments : JoVE
|October 14, 2024
概括
相对光和电子显微镜可视化了动态有机体相互作用. 这项研究揭示了 mitochondrial-lysosomal 接触在压力下如何发生变化,为细胞功能和病理学提供了洞察力.
科学领域:
- 细胞生物学 细胞生物学
- 显微镜技术 显微镜技术
背景情况:
- 细胞器官,如线粒体和溶解体,表现出动态结构,对细胞功能至关重要.
- 虽然传输电子显微镜提供了高分辨率,但光显微镜对于通过特定标签可视化动态有机细胞行为至关重要.
研究的目的:
- 为观察线粒体-溶解体相互作用提供双色相关光和电子显微镜 (CLEM) 协议.
- 研究这些相互作用如何在特定的细胞应激条件下发生变化.
主要方法:
- 线粒体被标记为mEosEM (线粒体-线粒体).
- 溶解体被标记为TMEM192-V5-APEX2.2. 这样一个标签.
- 采用双色CLEM可视化固定细胞中的器官相互作用.
主要成果:
- 巴菲洛米辛治疗 (抑制溶酶体功能) 增加了线粒体-溶酶体接触,导致溶酶体内的线粒体碎片化.
- U18666A治疗 (抑制溶酶体胆固醇出口) 导致溶酶体被线粒体包围.
- 在不同的压力条件下,CLEM成像揭示了不同的相互作用模式.
结论:
- 开发的双色CLEM方法有效地可视化了固定细胞中的线粒体-溶解体相互作用.
- 这种技术为研究器官活力及其在细胞病理学中的作用提供了强大的工具.
- 了解这些相互作用对于研究细胞功能和疾病机制至关重要.
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