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TitleMulti-scale 3D Cryo-Correlative Microscopy for Vitrified Cells.
Journal, issue, pagesStructure, Vol. 28, Issue 11, Page 1231-11237.e3, Year 2020
Publish dateNov 3, 2020
AuthorsGong-Her Wu / Patrick G Mitchell / Jesus G Galaz-Montoya / Corey W Hecksel / Emily M Sontag / Vimal Gangadharan / Jeffrey Marshman / David Mankus / Margaret E Bisher / Abigail K R Lytton-Jean / Judith Frydman / Kirk Czymmek / Wah Chiu /
PubMed AbstractThree-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 ...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.
External linksStructure / PubMed:32814034 / PubMed Central
MethodsEM (subtomogram averaging) / EM (tomography)
Resolution21.0 Å
Structure data

EMDB-21952:
Subtomogram average of ribosomes from tomograms of cryoFIB-SEM-generated lamellae of yeast cells under heat shock stress.
Method: EM (subtomogram averaging) / Resolution: 21.0 Å

EMDB-21953:
Downsampled and filtered tomogram of a region from a cryoFIB-SEM-generated lamellae of yeast cells under heat shock stress showing large protein aggregates
Method: EM (tomography)

Source
  • Saccharomyces cerevisiae (brewer's yeast)

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