在此之后,嵌入相对光和电子显微镜.
Shuyuan Wang1, Haiyan Xiong2, Qiyuan Chang3
1Public Technology Service Center, Fujian Medical University.
Journal of visualized experiments : JoVE
|January 29, 2024
概括
这项研究引入了Epon嵌入后相关光和电子显微镜 (CLEM) 的新协议. 它成功地保留了光和超结构,克服了CLEM技术之前的局限性.
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:
- 相对光和电子显微镜 (CLEM) 集成了光显微镜 (FM) 和电子显微镜 (EM) 进行细胞分析.
- 传统的CLEM通常涉及光信号和超结构保存之间的妥协.
- 与其他嵌入式介质相比,Epon树脂提供了优越的超结构保护.
研究的目的:
- 开发和详细说明EPON后嵌入CLEM的协议.
- 为了在CLEM中实现光和超结构的同时保存.
- 在CLEM中改进细胞识别和图像注册.
主要方法:
- 使用mEosEM,一种光蛋白被证明可以抵抗氧化固定和Epon嵌入.
- 实施详细的 EM 样品准备,FM 成像,EM 成像和图像对齐的详细步骤程序.
- 完善在FM和EM成像模式中识别相同细胞的方法,并改进图像注册.
主要成果:
- 成功的Epon后嵌入CLEM首次实现,同时保持光和超结构.
- 在氧化固定和Epon嵌入条件下证明了mEosEM的可行性.
- 改进了用于准确关联FM和EM数据的程序.
结论:
- 开发的协议可以同时进行高分辨率的超结构和光成像.
- 这种方法克服了CLEM以前的局限性,为成像模式提供了平衡的方法.
- 该协议旨在在标准电子显微镜设备中轻松实现.
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