使用相对 AFM/光显微镜成像核包裹
Emilie Costes1, Anthony Vial2, Christine Doucet3
1Centre de Biologie Structurale, INSERM, CNRS, University of Montpellier, Montpellier, France.
Methods in molecular biology (Clifton, N.J.)
|August 20, 2025
概括
这项研究提出了一种新方法,用于从哺乳动物细胞中制备核包裹 (NE),保持核孔复合体 (NPC) 结构. 该技术允许使用原子力显微镜 (AFM) 对内部和外部核膜进行高分辨率成像.
科学领域:
- 细胞生物学
- 生物物理
- 显微镜
背景情况:
- 核膜 (NE) 是一个具有内外膜的关键真核结构.
- 由于膜张力损失,现有的NE制备方法可以改变核孔综合体 (NPC) 形态.
- 原子力显微镜 (AFM) 是用于成像NE结构的强大工具.
研究的目的:
- 从培养的哺乳动物细胞中制备完整的核包裹 (NE) 的新方法.
- 在样本制备过程中保持核孔复合物的原生结构.
- 使用AFM和相关显微镜实现内外核膜的高分辨率成像.
主要方法:
- 从培养的哺乳动物细胞中提取细胞核并将其贴在玻璃基板上.
- 温和地打开核以保持膜张力和NPC完整性.
- 免疫标记结合AFM和直接的随机光学重建显微镜 (dSTORM) 进行相关成像.
主要成果:
- 这种新的制备方法成功地保持了NE膜张力和NPC结构.
- 实现内部和外部核膜的高分辨率AFM成像.
- 相对的AFM/dSTORM成像提供了详细的结构和分子信息.
结论:
- 该协议为研究具有保存形态的NE和NPC提供了一个强大的方法.
- 该方法适用于各种细胞模型,包括植物细胞和组织.
- 它可以提供先进的相关成像技术以获得更深入的生物学见解.
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