通过相关活细胞和体积电子显微镜对内溶酶体区的功能性表征
Nalan Liv1, Job Fermie2, Corlinda B M Ten Brink1
1Center for Molecular Medicine, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Methods in cell biology
|July 14, 2023
概括
这项研究引入了功能相关显微镜 (功能CLEM),将活细胞光生物传感器数据与高分辨率电子显微镜相结合. 这种新的方法揭示了详细的有机体功能和超结构,推进了细胞生物学研究.
科学领域:
- 细胞生物学 细胞生物学
- 显微镜的使用方法
- 生物物理学的生物物理.
背景情况:
- 光显微镜 (FM) 提供了对活细胞蛋白活性的洞察力,但缺乏超结构细节.
- 电子显微镜 (EM) 提供超结构上下文,但不是动态功能信息.
- 整合FM和EM对于理解细胞器官组织至关重要.
研究的目的:
- 开发一种将活细胞功能数据与高分辨率超结构集成的方法.
- 为了能够对细胞器官中的功能-超结构相关性进行定量分析.
- 为了研究晚期内溶酶体区的功能参数.
主要方法:
- 功能相关显微镜 (功能CLEM) 的发展.
- 用光生物传感器标记活细胞,然后进行电磁处理.
- 使用体积电子显微镜进行成像并将光信号与超结构相关联.
主要成果:
- 在单个数据集中成功关联了数百个光斑.
- 对内溶酶体区的功能-超结构数据进行定量分析.
- 揭示了酶活性和晚期内分泌体中高水平的显著差异.
结论:
- 功能性CLEM弥合了活细胞功能成像和超结构分析之间的差距.
- 该方法与各种探头兼容,可用于广泛的应用.
- 为各种细胞结构的大规模功能研究铺平了道路.
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