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通过亲和捕捉冷电子断层扫描揭示的细胞外丝

Leeya Engel^1,2, Magda Zaoralová³, Momei Zhou⁴

¹Dept. of Chemical Engineering, Stanford University, Stanford, CA, USA 94305.

bioRxiv : the preprint server for biology

|August 14, 2023

概括

这项研究提出了一种亲和性捕获方法,用于准备像T细胞这样的最小粘附细胞,用于高分辨率冷电子断层扫描 (cryo-ET) 成像. 该工作流提高了效率,并使细胞结构的纳米尺度可视化成为可能.

科学领域:

细胞和分子成像技术
生物物理学的生物物理.
显微镜技术显微镜技术

背景情况:

低温电子断层扫描 (cryo-ET) 为细胞架构提供纳米尺度的洞察力.
微纹对于为冷-ET工作流程准备附着细胞至关重要.
准备最小粘附细胞用于冷ET存在挑战.

研究的目的:

开发和演示微模式工作流程,用于使用亲和力捕获的最小附着细胞类型.
为了实现T细胞和Jurkat细胞的高分辨率冷电子断层扫描 (cryo-ET) 和冷聚焦离子束 (cryo-FIB) 成像.
提高工作流程效率和细胞定位,以获取冷ET数据.

主要方法:

开发用于细胞微模式的亲和力捕获系统.
将工作流应用于人类T细胞和Jurkat细胞.
集成与冷聚焦离子束 (cryo-FIB) 研磨和冷电子断层扫描 (cryo-ET) 数据采集.

主要成果:

成功的纳米级成像Jurkat细胞,揭示了细胞外丝状结构.
证明了工作流效率的提高,并始终生产定位良好的电池.
验证了对最小附着细胞类型的冷ET的亲和力捕获的实用性.

结论:

开发的亲和力捕获微模式工作流程有效地为冷ET准备了最小粘附细胞.
这种方法增强了用于研究细胞纳米级架构的成像能力.
该方法适用于各种附着和非附着细胞类型的高分辨率成像.

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