A mammalian system for high-resolution imaging of intact cells by cryo-electron tomography
| Autor: | Xia Li, Hangjun Wu, Yunjie Chang, Pei Wang, Jun Liu, Donghyun Park, Abhijith Radhakrishnan |
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| Rok vydání: | 2021 |
| Předmět: |
Organelles
Materials science Cryoelectron Microscopy Optical Imaging Biophysics High resolution Molecular resolution PC12 Cells Molecular machine Rats Imaging Three-Dimensional Molecular level Correlative light and electron microscopy Organelle Image Processing Computer-Assisted Animals Humans Cryo-electron tomography Molecular Biology High resolution imaging |
| Zdroj: | Progress in Biophysics and Molecular Biology. 160:87-96 |
| ISSN: | 0079-6107 |
| DOI: | 10.1016/j.pbiomolbio.2020.09.005 |
| Popis: | Mammalian cells contain an elaborate network of organelles and molecular machines that orchestrate essential cellular processes. Visualization of this network at a molecular level is vital for understanding these cellular processes. Here we present a model system based on nerve growth factor (NGF)-differentiated PC12 cells (PC12+) and suitable for high resolution imaging of organelles and molecular machines in situ. We detail an optimized imaging pipeline that effectively combines correlative light and electron microscopy (CLEM), cryo-focused ion beam (cryo-FIB), cryo-electron tomography (cryo-ET), and sub-tomogram averaging to produce three-dimensional and molecular resolution snapshots of organelles and molecular machines in near-native cellular environments. Our studies demonstrate that cryo-ET imaging of PC12+ systems provides an accessible and highly efficient avenue for dissecting specific cellular processes in mammalian cells at high resolution. |
| Databáze: | OpenAIRE |
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