TY - JOUR T1 - Correlative three-dimensional super-resolution and block face electron microscopy of whole vitreously frozen cells JF - bioRxiv DO - 10.1101/773986 SP - 773986 AU - David P. Hoffman AU - Gleb Shtengel AU - C. Shan Xu AU - Kirby R. Campbell AU - Melanie Freeman AU - Lei Wang AU - Daniel E. Milkie AU - H. Amalia Pasolli AU - Nirmala Iyer AU - John A. Bogovic AU - Daniel R. Stabley AU - Abbas Shirinifard AU - Song Pang AU - David Peale AU - Kathy Schaefer AU - Wim Pomp AU - Chi-Lun Chang AU - Jennifer Lippincott-Schwartz AU - Tom Kirchhausen AU - David J. Solecki AU - Eric Betzig AU - Harald Hess Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/09/18/773986.abstract N2 - Living cells function through the spatial compartmentalization of thousands of distinct proteins serving a multitude of diverse biochemical needs. Correlative super-resolution (SR) fluorescence and electron microscopy (EM) has emerged as a pathway to directly view nanoscale protein relationships to the underlying global ultrastructure, but has traditionally suffered from tradeoffs of structure preservation, fluorescence retention, resolution, and field of view. We developed a platform for three-dimensional correlative cryogenic SR and focused ion beam milled block-face EM across entire vitreously frozen cells that addresses these issues by preserving native ultrastructure and enabling independent SR and EM workflow optimization. Application to a variety of biological systems revealed a number of unexpected protein-ultrastructure relationships and underscored the value of a comprehensive multimodal view of ultrastructural variability across whole cells. ER -