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Article Dans Une Revue Research Square - Preprint Année : 2023

Facile hermetic TEM grid preparation for molecular imaging of hydrated biological samples at room temperature

Résumé

Abstract Although structures of vitrified supramolecular complexes have been determined at near-atomic resolution, elucidating in situ molecular structure in living cells remains a major challenge. Here, we apply a novel but simple liquid-cell technique, developed previously for real-time imaging of the dynamics at a liquid-gas interface, to image wet biological samples. With extra scattering from the liquid phase, the transmission electron micrographs show amplitude contrast comparable to that in negatively stained samples. Single-molecule domains are resolved in the protein complex GroEL imaged in buffer solution at room temperature. Moreover, various stages of virus cell entry, which are transient events with very few structural information to date, are also captured. Morphological details are reconstructed using the technique of individual particle electron tomography. These results demonstrate that this approach can be a valuable yet cost-effective technique complementary to other microscopy techniques for addressing important biological questions at the molecular level.

Dates et versions

hal-04384282 , version 1 (10-01-2024)

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Lingli Kong, Jianfang Liu, Meng Zhang, Zhuoyang Lu, Han Xue, et al.. Facile hermetic TEM grid preparation for molecular imaging of hydrated biological samples at room temperature. Research Square - Preprint, 2023, ⟨10.21203/rs.3.rs-2464569/v1⟩. ⟨hal-04384282⟩
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