National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
P01NS092525
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
5P41GM103832
United States
National Institutes of Health/National Institute on Aging (NIH/NIA)
PO1AG054407
United States
National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
F32NS086253
United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
S10OD021600
United States
Citation
Journal: Structure / Year: 2020 Title: Multi-scale 3D Cryo-Correlative Microscopy for Vitrified Cells. Authors: Gong-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- ...Authors: Gong-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 / 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 ...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.
History
Deposition
May 10, 2020
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Header (metadata) release
Sep 2, 2020
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Map release
Sep 2, 2020
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Update
Sep 2, 2020
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Current status
Sep 2, 2020
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
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