EMD-0733

Tomography
EMD-0733 Deposition: 07/08/2019
Map released: 12/08/2020
Last modified: 21/07/2021
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EMD-0733

Cryo electron tomogram of cryo-lamella of spinach leaf

EMD-0733

Tomography
EMD-0733 Deposition: 07/08/2019
Map released: 12/08/2020
Last modified: 21/07/2021
Overview Sample Experiment Validation Volume Browser Additional data Links
Sample Organism: Spinacia oleracea
Sample: cryo-lamella of spinach leaf
Raw data: EMPIAR-10302

Deposition Authors: Zhang J, Zhang D, Sun L, Sun F
VHUT-cryo-FIB, a method to fabricate frozen hydrated lamellae from tissue specimens for in situ cryo-electron tomography.
Zhang J, Zhang D, Sun L, Ji G, Huang X , Niu T, Xu J, Ma C, Zhu Y, Gao N , Xu W, Sun F
(2021) J Struct Biol , 213 , 107763 - 107763
PUBMED: 34174447
DOI: doi:10.1016/j.jsb.2021.107763
ISSN: 1095-8657
ASTM: JSBIEM
Abstract:
Cryo-electron tomography (cryo-ET) provides a promising approach to study intact structures of macromolecules in situ, but the efficient preparation of high-quality cryosections represents a bottleneck. Although cryo-focused ion beam (cryo-FIB) milling has emerged for large and flat cryo-lamella preparation, its application to tissue specimens remains challenging. Here, we report an integrated workflow, VHUT-cryo-FIB, for efficiently preparing frozen hydrated tissue lamella that can be readily used in subsequent cryo-ET studies. The workflow includes vibratome slicing, high-pressure freezing, ultramicrotome cryo-trimming and cryo-FIB milling. Two strategies were developed for loading cryo-lamella via a side-entry cryo-holder or an FEI AutoGrid. The workflow was validated by using various tissue specimens, including rat skeletal muscle, rat liver and spinach leaf specimens, and in situ structures of ribosomes were obtained at nanometer resolution from the spinach and liver samples.