冷冻聚焦离子束-扫描电镜成像技术研究进展

Advances in Cryo-focused Ion Beam-Scanning Electron Microscopy Imaging Technology

冷冻聚焦离子束-扫描电镜成像(Cryo-FIB-SEM)是一种新兴的成像检测技术,在原位进行冷冻聚焦离子束切割和冷冻扫描电镜成像,为研究天然含水状态下生物样品内部未被破坏的原始结构打开了一扇窗口。近年来,该技术在生命科学领域的应用研究取得了一系列重要进展。该文对其在冷冻体积连续成像、冷冻光电关联成像、冷冻透射扫描成像、冷冻含水切片制备监控及冷冻扫描图像处理等方面的研究进展进行综述,并展望了该技术在大体积生物样品三维原位成像研究领域的前沿性发展趋势,以期推动Cryo-FIB-SEM技术在生物样品三维结构研究中的应用。

Cryo-focused ion beam-scanning electron microscopy(Cryo-FIB-SEM) is an emerging technology designed for advanced imaging detection, which performs in situ by combining Cryo-FIB milling and Cryo-SEM imaging, and has facilitated the visualization of the native structures of biological sample in the context of the cellular environment in the frozen hydrated state. In recent years, a series of important advances have been achieved in the application of this technology in the research field of life science. In this review, we summarize its application in cryo-volume serial imaging,and in combination with cryo-correlative light and electron microscopy(CLEM), cryo-transmission SEM(TSEM),cryo-lamella preparation monitoring, and Cryo-SEM image processing. We also provide future prospective on future development and application of Cryo-FIB-SEM in three-dimensional in situ imaging of large volume biological samples.