CryoEM has become the method of choice for determining the structure of large macromolecular complexes in multiple conformations, at resolutions where unambiguous atomic models can be built. Two effects that have limited progress in single-particle cryoEM are (i) beam-induced movement during image acquisition and (ii) protein adsorption and denaturation at the air-water interface during specimen preparation. While beam-induced movement now appears to have been resolved by all-gold specimen support grids with very small holes, surface effects at the air-water interface are a persistent problem. Strategies to overcome these effects include the use of alternative support films and new techniques for specimen deposition. We examine the future potential of recording perfect images of biological samples for routine structure determination at atomic resolution.

中文翻译：

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单粒子cryoEM对高分辨率结构测定的当前限制

CryoEM 已成为确定多种构象的大分子复合物结构的首选方法，其分辨率可以建立明确的原子模型。限制单粒子 cryoEM 进展的两个影响是 (i) 图像采集过程中的光束诱导运动和 (ii) 样品制备过程中空气-水界面处的蛋白质吸附和变性。虽然现在似乎已经通过带有非常小孔的全金试样支撑网格解决了梁引起的运动，但空气-水界面的表面效应是一个长期存在的问题。克服这些影响的策略包括使用替代支撑膜和用于样品沉积的新技术。