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高精度无抗体微管标记用于膨胀显微镜

Rajdeep Chowdhury^1,2, Donatus Krah¹, Antonios Ntolkeras¹

¹Department of Neuro- and Sensory Physiology, University Medical Center Göttingen, Göttingen, Germany.

|December 26, 2025

概括

本研究介绍了一种优化的扩展显微镜 (ExM) 协议,用于精确的微管可视化. 该方法使用洗剂提取和NHS-标签用于高分辨率纳米尺度成像,提高生物样本的准确性.

科学领域:

细胞生物学细胞生物学
显微镜技术显微镜技术
生物化学生物化学

背景情况:

扩展显微镜 (ExM) 提供纳米级成像,但在准确标记细胞骨纤维如微管中面临挑战.
现有方法在样本准备过程中可能会遭受结构扭曲和标签不准确.

研究的目的:

使用ExM开发一个优化的协议,用于高精度的微管可视化.
提高微管结构纳米尺度成像的准确性和可靠性.

主要方法:

结合洗剂提取来隔离微管和去除细胞碎片.
使用NHS-ester标记N-终端在消化的素上,避免抗体.
集成到现有的ExM工作流程中,用于体外和细胞样本.

主要成果:

实现细胞质成分和膜的选择性去除,保持微管的超结构.
允许精确的分子定位,最小的链接误差和高信号噪声比 (SNR).
在纳米尺度分辨率上展示了有效的光体移位,这取决于膨胀因子.

结论:

优化的协议为ExM中高精度微管体成像提供了强大的方法.
被标记的微管可以作为内部参考标准来纠正ExM数据集中的扩张因子.
这种技术增强了ExM在纳米尺度上研究细胞骨组织的实用性.

关键词:

洗剂提取的方法扩展显微镜扩展显微镜高精度的成像成像技术这些微管是微管子.在NHS-ester标签上

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相关概念视频

Microtubules

Microtubules

There are three types of cytoskeletal structures in eukaryotic cells—microfilaments, intermediate filaments, and microtubules. With a diameter of about 25 nm, microtubules are the thickest of these fibers. Microtubules carry out a variety of functions that include cell structure and support, transport of organelles, cell motility (movement), and the separation of chromosomes during cell division.