Characterizing the resolution and throughput of the Apollo direct electron detector.
Characterizing the resolution and throughput of the Apollo direct electron detector.
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表征 Apollo 直接电子探测器的分辨率和吞吐量。
DOI：
10.1016/j.yjsbx.2022.100080
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发表时间：
2023
影响因子：
2.9
通讯作者：
Stagg, Scott M.
中科院分区：
文献类型：
--
作者：
Peng, Ruizhi;Fu, Xiaofeng;Mendez, Joshua H.;Randolph, Peter S.;Bammes, Benjamin E.;Stagg, Scott M.
一种新的高分辨率直接电子探测相机，阿波罗，其特点。Apollo使用实时计数来增加冷冻EM的数据采集吞吐量。Apollo可以在高达60 eps的剂量率下产生优于1.9 μ s分辨率的重建。电子探测技术的进步对高分辨率冷冻电镜结构测定的成功至关重要。Direct Electron公司开发了新一代的直接电子探测器Apollo。Apollo采用了一种新型的基于事件的MAPS探测器，专为超快电子计数而设计。我们对这款新相机进行了评估，发现它具有高量子探测效率（DQE）和低符合损失，能够以每秒每像素近80个输入电子的速度采集高质量的电子计数数据。我们进一步表征了Apollo在真实的生物样品上用于单粒子冷冻EM的性能。使用小鼠脱铁铁蛋白，Apollo在所有三种测试剂量率下从每个半天的数据收集会话中获得了优于1.9 μ m分辨率的重建。通过延长样品采集时间和改进样品制备方法，将小鼠脱铁铁蛋白的分辨率重建到1.66 μ m。应用于更具挑战性的小蛋白质醛缩酶，我们获得了2.24 μ m分辨率的重建。高质量的图谱表明Apollo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，并能够捕获高质量的数据，产生高分辨率的重建大小单粒子样品。
A new high-resolution direct electron detection camera, the Apollo, is characterized. The Apollo uses on-the-fly counting to increase data acquisition throughput for cryo-EM. The Apollo can produce better than 1.9 Å resolution reconstructions at dose rates up to 60 eps. 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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影响因子：
3.9
作者：
Cash JN;Kearns S;Li Y;Cianfrocco MA
通讯作者：
Cianfrocco MA
影响因子：
64.8
作者：
Nakane T;Kotecha A;Sente A;McMullan G;Masiulis S;Brown PMGE;Grigoras IT;Malinauskaite L;Malinauskas T;Miehling J;Uchański T;Yu L;Karia D;Pechnikova EV;de Jong E;Keizer J;Bischoff M;McCormack J;Tiemeijer P;Hardwick SW;Chirgadze DY;Murshudov G;Aricescu AR;Scheres SHW
通讯作者：
Scheres SHW
影响因子：
2.2
作者：
McMullan, G.;Chen, S.;Henderson, R.;Faruqi, A. R.
通讯作者：
Faruqi, A. R.
影响因子：
3
作者：
Glaeser RM;Typke D;Tiemeijer PC;Pulokas J;Cheng A
通讯作者：
Cheng A
DOI：
10.1107/s2059798319011471
发表时间：
2019-10-01
影响因子：
2.2
作者：
Liebschner, Dorothee;Afonine, Pavel V.;Adams, Paul D.
通讯作者：
Adams, Paul D.