In situ cryo-electron tomography: a new method to elucidate cytoplasmic zoning at the molecular level

• Published in: Journal of biochemistry (Tokyo) Vol. 175; no. 2; pp. 187 - 193
• Main Authors: Chen, Lin, Fukata, Yuko, Murata, Kazuyoshi
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 25.02.2024
• Subjects: Aqueous solutions; Biomolecules; Electron microscopy; Ion beams; Liquid phases; Phase separation; Scanning electron microscopy; Structural analysis; Tomography; Zoning; cryo-electron tomography; liquid–liquid phase separation; cryo-focused ion beam scanning electron microscope; cytoplasmic zoning; cryo-electron microscopy; liquid-liquid phase separation
• ISSN: 0021-924X; 1756-2651; 1756-2651
• DOI: 10.1093/jb/mvad102

Abstract Cryo-electron microscopy was developed as a powerful tool for imaging biological specimens in near-native conditions. Nowadays, advances in technology, equipment and computations make it possible to obtain structures of biomolecules with near-atomic resolution. Furthermore, cryo-electron tomography combined with continuous specimen tilting allows structural analysis of heterogeneous biological specimens. In particular, when combined with a cryo-focused ion beam scanning electron microscope, it becomes possible to directly analyse the structure of the biomolecules within cells, a process known as in situ cryo-electron tomography. This technique has the potential to visualize cytoplasmic zoning, involving liquid–liquid phase separation, caused by biomolecular networks in aqueous solutions, which has been the subject of recent debate. Here, we review advances in structural studies of biomolecules to study cytoplasmic zoning by in situ cryo-electron tomography. Graphical Abstract Graphical Abstract