RT Journal Article
SR Electronic
T1 Multi-Scale 3D Cryo-Correlative Microscopy for Vitrified Cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.05.21.107771
DO 10.1101/2020.05.21.107771
A1 Gong-Her Wu
A1 Patrick G. Mitchell
A1 Jesus G. Galaz-Montoya
A1 Corey W. Hecksel
A1 Emily M. Sontag
A1 Vimal Gangadharan
A1 Jeffrey Marshman
A1 David Mankus
A1 Margaret E. Bisher
A1 Abigail K. R. Lytton-Jean
A1 Judith Frydman
A1 Kirk Czymmek
A1 Wah Chiu
YR 2020
UL http://biorxiv.org/content/early/2020/05/22/2020.05.21.107771.abstract
AB 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-labelled 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.Competing Interest StatementVimal Gangadharan and Jeffrey Marshman work for Zeiss, the developer of the cryo-Airyscan confocal microscope, one of the instruments used in this study. All other authors have no conflicts of interest to declare.