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

Koch Institute for Integrative Cancer Research

Massachusetts Institute of Technology

CryoEM Status Update and Innovative Developments

Margaret E. Bisher

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Koch Institute for Integrative Cancer Research

Massachusetts Institute of Technology

CryoEM Status Update and Innovative Developments

Koch Institute for Integrative Cancer Research