Premaraj Navya, Heeren Ron M A, Ravelli Raimond B G, Knoops Kèvin
Maastricht MultiModal Molecular Imaging Institute(M4I), Maastricht University, Maastricht 6229 ER, The Netherlands.
Microscopy CORE Lab, Maastricht University, Maastricht 6229 ER, The Netherlands.
Anal Chem. 2025 Jun 17;97(23):11959-11967. doi: 10.1021/acs.analchem.5c01534. Epub 2025 Jun 6.
Cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET) have revolutionized structural biology by enabling high-resolution imaging of biomolecules and cellular structures. However, traditional sample carriers, such as copper grids with carbon films, presented limitations, particularly in cryo-ET workflows. Issues like uneven cell distribution, beam-induced motion, and suboptimal vitrification can compromise data quality. Recent advances in sample carrier design have successfully addressed key challenges, including the development of gold-based supports, graphene coatings, and nanofluidic chips. These innovations have improved mechanical stability, enhanced thermal conductivity, and provided better control over ice layer uniformity, leading to more consistent sample preparation and higher-quality imaging. In this perspective, sample preparation advancements, novel approaches such as titanium autogrids and slot grids with continuous gold foils, and their role toward future cryo-ET applications are discussed. These new designs have the potential to simplify workflows and optimize cell growth environments. Furthermore, this perspective highlights how integrating these cutting-edge technologies with prior advancements in sample carrier design can enhance cryo-ET workflows. Combined, they enable cryo-EM imaging of thicker samples and drive progress in structural biology research.
冷冻电子显微镜（cryo-EM）和冷冻电子断层扫描（cryo-ET）通过实现对生物分子和细胞结构的高分辨率成像，彻底改变了结构生物学。然而，传统的样品载体，如带有碳膜的铜网，存在局限性，特别是在冷冻电子断层扫描工作流程中。细胞分布不均、束流诱导运动和玻璃化效果欠佳等问题会影响数据质量。样品载体设计的最新进展成功解决了关键挑战，包括开发基于金的支撑体、石墨烯涂层和纳米流体芯片。这些创新提高了机械稳定性，增强了热导率，并能更好地控制冰层均匀性，从而实现更一致的样品制备和更高质量的成像。本文探讨了样品制备的进展、诸如带有连续金箔的钛自动网格和狭缝网格等新方法及其在未来冷冻电子断层扫描应用中的作用。这些新设计有可能简化工作流程并优化细胞生长环境。此外，本文强调了将这些前沿技术与样品载体设计的先前进展相结合如何能够增强冷冻电子断层扫描工作流程。综合起来，它们能够对更厚的样品进行冷冻电子显微镜成像，并推动结构生物学研究取得进展。
