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

Lingli Kong, Jianfang Liu, Meng Zhang, Zhuoyang Lu, Han Xue, Amy Ren, Jiankang Liu, Jinping Li, Wai Li Ling () and Gang Ren ()
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Lingli Kong: Lawrence Berkeley National Laboratory
Jianfang Liu: Lawrence Berkeley National Laboratory
Meng Zhang: Lawrence Berkeley National Laboratory
Zhuoyang Lu: Xi’an Jiaotong University
Han Xue: Lawrence Berkeley National Laboratory
Amy Ren: University of California
Jiankang Liu: Xi’an Jiaotong University
Jinping Li: Mayo Clinic
Wai Li Ling: Université Grenoble Alpes, CEA, CNRS, IBS
Gang Ren: Lawrence Berkeley National Laboratory

Nature Communications, 2023, vol. 14, issue 1, 1-10

Abstract: 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 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.

Date: 2023
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DOI: 10.1038/s41467-023-41266-x

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