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-Structure paper
Title | Facile hermetic TEM grid preparation for molecular imaging of hydrated biological samples at room temperature. |
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Journal, issue, pages | Nat Commun, Vol. 14, Issue 1, Page 5641, Year 2023 |
Publish date | Sep 13, 2023 |
Authors | Lingli Kong / Jianfang Liu / Meng Zhang / Zhuoyang Lu / Han Xue / Amy Ren / Jiankang Liu / Jinping Li / Wai Li Ling / Gang Ren / |
PubMed Abstract | Although structures of vitrified supramolecular complexes have been determined at near-atomic resolution, elucidating in situ molecular structure in living cells remains a challenge. Here, we report ...Although structures of vitrified supramolecular complexes have been determined at near-atomic resolution, elucidating in situ molecular structure in living cells remains a challenge. Here, we report a straightforward liquid cell technique, originally developed for real-time visualization of dynamics at a liquid-gas interface using transmission electron microscopy, to image wet biological samples. Due to the scattering effects from the liquid phase, the micrographs display an amplitude contrast comparable to that observed in negatively stained samples. We succeed in resolving subunits within the protein complex GroEL imaged in a buffer solution at room temperature. Additionally, we capture various stages of virus cell entry, a process for which only sparse structural data exists due to their transient nature. To scrutinize the morphological details further, we used individual particle electron tomography for 3D reconstruction of each virus. These findings showcase this approach potential as an efficient, cost-effective complement to other microscopy technique in addressing biological questions at the molecular level. |
External links | Nat Commun / PubMed:37704637 / PubMed Central |
Methods | EM (tomography) |
Resolution | 65.0 - 74.0 Å |
Structure data | EMDB-25890: Single-molecule 3D density map of HIV cellular entry by liquid-phase electron tomography (particle #1) EMDB-25894: Single-molecule 3D density map of HIV cellular entry by liquid-phase electron tomography (particle #3) EMDB-25895: Single-molecule 3D density map of HIV cellular entry by liquid-phase electron tomography (particle #2) |
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