一步4x和12x3D-ExM:在细胞生物学中强大的超分辨率显微镜
Roshan X Norman1,2,3, Yu-Chia Chen2,4, Emma E Recchia2
1Biophysics Graduate Program, University of Wisconsin-Madison, Madison, Wisconsin.
bioRxiv : the preprint server for biology
|August 26, 2024
概括
扩展显微镜 (ExM) 为细胞生物学提供先进的超高分辨率成像. 优化的4x和12x3D-ExM方法提供了成本效益高,用户友好的细胞结构纳米尺度可视化.
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:
- 超分辨率显微镜为生物成像提供纳米级分辨率.
- 扩展显微镜 (ExM) 与电子显微镜等传统方法相比具有优势.
- ExM 能够同时进行标签和更简单的样品准备.
研究的目的:
- 介绍优化的4倍和12倍的3D-同位素和保存扩展显微镜 (4x和12x3D-ExM) 方法.
- 展示3D-ExM在细胞生物学研究中用于高分辨率成像的实用性.
主要方法:
- 开发和优化了4x和12x的3D-ExM协议.
- 使用标准的共聚焦显微镜来成像扩展样本.
- 对二维和三维细胞培养模型的应用方法.
主要成果:
- 实现了强大的和可重复的3D同位素扩张.
- 12x 3D-ExM在标准共聚焦显微镜下产生了30nm以下的侧面分辨率.
- 启用了纳米级结构的可视化,如染色体,kinetochores和病毒粒子.
结论:
- 3D-ExM是一种具有成本效益和用户友好的超高分辨率技术.
- 优化的3D-ExM方法适用于各种细胞生物学应用.
- 促进了纳米级细胞和染色质结构的详细研究.
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