Cryo-electron microscopy (cryoEM) has become one of the most important approach for structural biology. However, barriers are still there for an increased successful rate, a better resolution and improved efficiency from sample preparation, data collection to image processing. CryoEM sample preparation is one of the bottlenecks with many efforts made recently, including the optimization of supporting substrate (e.g. ultra-thin carbon, graphene, pure gold, 2d crystal of streptavidin, and affinity modification), which was aimed to solve air-water interface problem, or reduce beam induced motion (BIM), or change particle distribution in the grid hole. Here, we report another effort of developing a new supporting substrate, the amorphous nickel-titanium alloy (ANTA) film, for cryoEM sample preparation as a layer of holey supporting film covering on TEM grid. Our investigations showed advantages of ANTA film in comparison with conventional carbon film, including much better electron conductivity and trace non-specific interaction with protein. These advantages yield less BIM and significantly improved particle distribution during cryoEM experiment of human apo-ferritn, thus resulting an improved reconstruction resolution from a reduced number of micrographs and particles. Unlike the pure gold film, the usage of the ANTA film is just same with the carbon film, compatible to conventional automatic cryoEM data collection procedure.

冷冻电子显微镜 (cryoEM) 已成为结构生物学最重要的方法之一。然而，从样品制备、数据收集到图像处理，在提高成功率、更好的分辨率和提高效率方面仍然存在障碍。CryoEM 样品制备是近期许多努力的瓶颈之一，包括优化支撑基底（例如超薄碳、石墨烯、纯金、链霉亲和素的二维晶体、亲和修饰），旨在解决空气-水界面问题，或减少光束诱导运动 (BIM)，或改变网格孔中的粒子分布。在这里，我们报告了开发新支撑基板的另一项努力，即非晶镍钛合金 (ANTA) 薄膜，用于冷冻电镜样品制备，作为覆盖在 TEM 网格上的一层多孔支撑膜。我们的研究表明安踏薄膜与传统碳薄膜相比具有优势，包括更好的电子传导性和与蛋白质的非特异性相互作用。这些优势在人类脱脂蛋白铁蛋白的冷冻电镜实验期间产生更少的 BIM 和显着改善的颗粒分布，从而通过减少的显微照片和颗粒数量提高重建分辨率。与纯金膜不同，ANTA 膜的使用与碳膜相同，兼容传统的自动冷冻电镜数据采集程序。包括更好的电子传导性和与蛋白质的非特异性相互作用。这些优势在人类脱脂蛋白铁蛋白的冷冻电镜实验期间产生更少的 BIM 和显着改善的颗粒分布，从而通过减少的显微照片和颗粒数量提高重建分辨率。与纯金膜不同，ANTA 膜的使用与碳膜相同，兼容传统的自动冷冻电镜数据采集程序。包括更好的电子传导性和与蛋白质的非特异性相互作用。这些优势在人类脱脂蛋白铁蛋白的冷冻电镜实验期间产生更少的 BIM 和显着改善的颗粒分布，从而通过减少的显微照片和颗粒数量提高重建分辨率。与纯金膜不同，ANTA 膜的使用与碳膜相同，兼容传统的自动冷冻电镜数据采集程序。