EMD-33910

Tomography
EMD-33910 Deposition: 26/07/2022
Map released: 09/11/2022
Last modified: 13/12/2023
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EMD-33910

In situ cryo-electron tomogram of collagen fibril in fibrosis liver tissue 1

EMD-33910

Tomography
EMD-33910 Deposition: 26/07/2022
Map released: 09/11/2022
Last modified: 13/12/2023
Overview Sample Experiment Validation Volume Browser Additional data Links
Sample Organism: Mus musculus
Sample: collagen fibril in fibrosis liver tissue

Deposition Authors: Wang S , Zhou H , Chen W , Jiang Y , Yan X, You H , Li X
CryoFIB milling large tissue samples for cryo-electron tomography.
Wang S , Zhou H , Chen W , Jiang Y , Yan X, You H , Li X
(2023) Sci Rep , 13 , 5879 - 5879
PUBMED: 37041258
DOI: doi:10.1038/s41598-023-32716-z
ISSN: 2045-2322
Abstract:
Cryo-electron tomography (cryoET) is a powerful tool for exploring the molecular structure of large organisms. However, technical challenges still limit cryoET applications on large samples. In particular, localization and cutting out objects of interest from a large tissue sample are still difficult steps. In this study, we report a sample thinning strategy and workflow for tissue samples based on cryo-focused ion beam (cryoFIB) milling. This workflow provides a full solution for isolating objects of interest by starting from a millimeter-sized tissue sample and ending with hundred-nanometer-thin lamellae. The workflow involves sample fixation, pre-sectioning, a two-step milling strategy, and localization of the object of interest using cellular secondary electron imaging (CSEI). The milling strategy consists of two steps, a coarse milling step to improve the milling efficiency, followed by a fine milling step. The two-step milling creates a furrow-ridge structure with an additional conductive Pt layer to reduce the beam-induced charging issue. CSEI is highlighted in the workflow, which provides on-the-fly localization during cryoFIB milling. Tests of the complete workflow were conducted to demonstrate the high efficiency and high feasibility of the proposed method.