表征阿波罗直接电子探测器的分辨率和吞吐量

IF 3.5 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Ruizhi Peng , Xiaofeng Fu , Joshua H. Mendez , Peter S. Randolph , Benjamin E. Bammes , Scott M. Stagg
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引用次数: 4

摘要

电子探测技术的进步对高分辨率低温电镜结构测定的成功至关重要。新一代的直接电子探测器被称为阿波罗,已经由直接电子公司开发。阿波罗使用了一种新颖的基于事件的map探测器,专为超快速电子计数而设计。我们对这款新相机进行了评估,发现它提供了高探测量子效率(DQE)和低重合损耗,能够以每秒每像素近80个输入电子的速度获得高质量的电子计数数据。我们进一步表征了Apollo在真实生物样品上的单粒子低温电镜性能。使用小鼠载铁蛋白,阿波罗在所有三种测试剂量率下获得了优于1.9 Å的分辨率重建。随着采集时间的延长和标本制备方法的改进,小鼠载铁蛋白重构至1.66 Å分辨率。应用于更具挑战性的小蛋白醛缩酶,我们获得了2.24 Å分辨率重建。高质量的地图表明阿波罗具有足够高的DQE,可以高保真地重建较小的蛋白质和复合物。我们的研究结果表明,阿波罗相机在广泛的剂量率范围内表现良好,能够捕获高质量的数据,为大小单颗粒样品产生高分辨率的重建。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Characterizing the resolution and throughput of the Apollo direct electron detector

Characterizing the resolution and throughput of the Apollo direct electron detector

Advances in electron detection have been essential to the success of high-resolution cryo-EM structure determination. A new generation of direct electron detector called the Apollo, has been developed by Direct Electron. The Apollo uses a novel event-based MAPS detector custom designed for ultra-fast electron counting. We have evaluated this new camera, finding that it delivers high detective quantum efficiency (DQE) and low coincidence loss, enabling high-quality electron counting data acquisition at up to nearly 80 input electrons per pixel per second. We further characterized the performance of Apollo for single particle cryo-EM on real biological samples. Using mouse apoferritin, Apollo yielded better than 1.9 Å resolution reconstructions at all three tested dose rates from a half-day data collection session each. With longer collection time and improved specimen preparation, mouse apoferritin was reconstructed to 1.66 Å resolution. Applied to a more challenging small protein aldolase, we obtained a 2.24 Å resolution reconstruction. The high quality of the map indicates that the Apollo has sufficiently high DQE to reconstruct smaller proteins and complexes with high-fidelity. Our results demonstrate that the Apollo camera performs well across a broad range of dose rates and is capable of capturing high quality data that produce high-resolution reconstructions for large and small single particle samples.

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来源期刊
Journal of Structural Biology: X
Journal of Structural Biology: X Biochemistry, Genetics and Molecular Biology-Structural Biology
CiteScore
6.50
自引率
0.00%
发文量
20
审稿时长
62 days
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