cryoem-status-update-and-innovative-developments

CryoEM Status Update and Innovative Developments

ARTICLES DISCUSSED:
Truong, C. D. et al. Sample preparation using a lipid monolayer method for electron crystallographic studies. Journal of Visualized ...

Massachusetts Institute of Technology

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.

Margaret E. Bisher

Biography

JoVE Journal Publications

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