气候变化领域集成服务门户(CCPortal): Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells

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DOI	10.1126/science.aaz5357
	Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells
	Hoffman D.P.; Shtengel G.; Xu C.S.; Campbell K.R.; Freeman M.; Wang L.; Milkie D.E.; Pasolli H.A.; Iyer N.; Bogovic J.A.; Stabley D.R.; Shirinifard A.; Pang S.; Peale D.; Schaefer K.; Pomp W.; Chang C.-L.; Lippincott-Schwartz J.; Kirchhausen T.; Solecki D.J.; Betzig E.; Hess H.F.
发表日期	2020
ISSN	0036-8075
卷号	367 期号:6475
英文摘要	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. © 2020 American Association for the Advancement of Science. All rights reserved.
关键词	cell compartmentalization cryopreservation electron microscopy image resolution protein three-dimensional modeling ultrastructure article chromatin electron microscopy endoplasmic reticulum genetic transcription human cell mammal cell nerve cell culture protein domain ultrastructure whole cell workflow animal cell adhesion cells Chlorocebus aethiops cryoelectron microscopy CV-1 cell line fluorescence microscopy freezing HeLa cell line human mouse procedures three-dimensional imaging tumor cell line ultrastructure Mammalia Animals Cell Adhesion Cell Line, Tumor Cells Chlorocebus aethiops COS Cells Cryoelectron Microscopy Freezing HeLa Cells Humans Imaging, Three-Dimensional Mice Microscopy, Fluorescence
语种	英语
来源机构	Science
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/133830
推荐引用方式 GB/T 7714	Hoffman D.P.,Shtengel G.,Xu C.S.,et al. Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells[J]. Science,2020,367(6475).
APA	Hoffman D.P..,Shtengel G..,Xu C.S..,Campbell K.R..,Freeman M..,...&Hess H.F..(2020).Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells.,367(6475).
MLA	Hoffman D.P.,et al."Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells".367.6475(2020).