Document Type

Article

Language

eng

Publication Date

11-3-2020

Publisher

Elsevier (Cell Press)

Source Publication

Structure

Source ISSN

0969-2126

Abstract

Three-dimensional (3D) visualization of vitrified cells can uncover structures of subcellular complexes without chemical fixation or staining. Here, we present a pipeline integrating three imaging modalities to visualize the same specimen at cryogenic temperature at different scales: cryo-fluorescence confocal microscopy, volume cryo-focused ion beam scanning electron microscopy, and transmission cryo-electron tomography. Our proof-of-concept benchmark revealed the 3D distribution of organelles and subcellular structures in whole heat-shocked yeast cells, including the ultrastructure of protein inclusions that recruit fluorescently-labeled chaperone Hsp104. Since our workflow efficiently integrates imaging at three different scales and can be applied to other types of cells, it could be used for large-scale phenotypic studies of frozen-hydrated specimens in a variety of healthy and diseased conditions with and without treatments.

Comments

Accepted version. Structure, Vol. 28, No. 11 (November 3, 2020): 1231-1237.e3. DOI. © 2020 Elsevier (Cell Press). Used with permission.

Emily M. Sontag was affiliated with Stanford University at the time of publication.

Available for download on Wednesday, November 03, 2021

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