| Literature DB >> 31949053 |
David P Hoffman1, Gleb Shtengel1, C Shan Xu1, Kirby R Campbell2, Melanie Freeman1, Lei Wang3,4,5, Daniel E Milkie1, H Amalia Pasolli1, Nirmala Iyer1, John A Bogovic1, Daniel R Stabley6, Abbas Shirinifard7, Song Pang1, David Peale1, Kathy Schaefer1, Wim Pomp3,4,5, Chi-Lun Chang1, Jennifer Lippincott-Schwartz1, Tom Kirchhausen1,3,4,5, David J Solecki2, Eric Betzig8,9,10,11,12,13, Harald F Hess8.
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.Entities:
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Year: 2020 PMID： 31949053 PMCID： PMC7339343 DOI： 10.1126/science.aaz5357
Source DB: PubMed Journal: Science ISSN： 0036-8075 Impact factor: 47.728