美国霍华德休斯医学研究所Harald F. Hess、Eric Betzig等研究人员合作取得一项新突破，他们开发出玻璃体冷冻细胞的三维超分辨率和电子相关显微镜。这一研究成果于2020年1月17日发表在国际学术期刊《科学》上。
研究人员为整个玻璃体冷冻细胞中的三维低温超分辨率（SR）和聚焦离子束电子显微镜（EM）开发了平台。该方法保留了超微结构，同时支持独立的SR和EM工作流程优化。研究人员在哺乳动物细胞中发现了意想不到的蛋白质-超微结构关系，包括含有内质网相关蛋白的核内囊泡、培养的神经元之间的网状粘附以及根据转录活性亚分类的染色质结构域。这些发现表明了整个细胞中超微结构变化的全面多模态视图的价值。
据介绍，在细胞内，数千种不同蛋白质的空间分隔可满足多种多样的生化需求。相关的SR荧光和EM可以阐明蛋白质与全局超微结构的空间关系，但在结构保留、荧光维持、分辨率和视野之间存在折衷。
附：英文原文
Title: Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells
Author: David P. Hoffman, Gleb Shtengel, C. Shan Xu, Kirby R. Campbell, Melanie Freeman, Lei Wang, Daniel E. Milkie, H. Amalia Pasolli, Nirmala Iyer, John A. Bogovic, Daniel R. Stabley, Abbas Shirinifard, Song Pang, David Peale, Kathy Schaefer, Wim Pomp, Chi-Lun Chang, Jennifer Lippincott-Schwartz, Tom Kirchhausen, David J. Solecki, Eric Betzig, Harald F. Hess
Issue&Volume: 2020/01/17
Abstract: Within cells, the spatial compartmentalization of thousands of distinct proteins serves a multitude of diverse biochemical needs. Correlative super-resolution (SR) fluorescence and electron microscopy (EM) can elucidate protein spatial relationships to global ultrastructure, but has suffered from tradeoffs of structure preservation, fluorescence retention, resolution, and field of view. We developed a platform for three-dimensional cryogenic SR and focused ion beam–milled block-face EM across entire vitreously frozen cells. The approach preserves ultrastructure while enabling independent SR and EM workflow optimization. We discovered unexpected protein-ultrastructure relationships in mammalian cells including intranuclear vesicles containing endoplasmic reticulum–associated proteins, web-like adhesions between cultured neurons, and chromatin domains subclassified on the basis of transcriptional activity. Our findings illustrate the value of a comprehensive multimodal view of ultrastructural variability across whole cells.
DOI: 10.1126/science.aaz5357
Source: https://science.sciencemag.org/content/367/6475/eaaz5357