Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells.

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-l

SUBMITTER: Hoffman DP 

PROVIDER: S-EPMC7339343 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature