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.
英文关键词	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
来源期刊	Science
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/244034
作者单位	Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, United States; Department of Developmental Neurobiology, St. Jude Children’s Research Hospital, Memphis, TN 38105, United States; Department of Cell Biology, Harvard Medical School, Boston, MA 02115, United States; Program in Cellular and Molecular Medicine, Boston Children’s Hospital, Boston, MA 02115, United States; Department of Pediatrics, Harvard Medical School, Boston, MA 02115, United States; Neuroimaging Laboratory, St. Jude Children’s Research Hospital, Memphis, TN 38105, United States; Bioimage Analysis Core, St. Jude Children’s Research Hospital, Memphis, TN 38105, United States; Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, United States; Department of Physics, University of California, Berkeley, CA 94720, United States; Howard Hughes Medical Institute, Berkeley, CA 94720, United States; Helen Wills Neuroscience Institute, Berkeley, CA 94720, United States; Molecula...
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],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.Science,367(6475).
MLA	Hoffman D.P.,et al."Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells".Science 367.6475(2020).