Rapid Tilt-Series Acquisition for Electron Cryotomography

Rapid Tilt-Series Acquisition for Electron Cryotomography
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用于电子冷冻断层扫描的快速倾斜系列采集

DOI:
10.1101/454587
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发表时间:
2018
期刊:
bioRxiv
影响因子:
--
通讯作者:
G. Jensen
G. Jensen
中科院分区:
--
文献类型:
--
作者:
G. Chreifi;Songye Chen;L. Metskas;M. Kaplan;G. Jensen

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使用配备单轴支架的新型Titan Krios平台,我们开发了两种加速倾斜系列收集的方法。我们展示了一种连续倾斜方法,可以在几秒钟内记录倾斜序列(比当前方法快100倍),但细节损失小于4纳米。我们还演示了一种快速增量方法,该方法可以以比当前方法快10倍的速度记录倾斜序列,并且具有相似的分辨率。我们描述了这两种方法在实际生物电子冷冻层析工作流程中的效用。我们确定了硬件和软件进一步改进的机会,并推测这些进步可能对结构生物学产生的影响。
Using a new Titan Krios stage equipped with a single-axis holder, we developed two methods to accelerate the collection of tilt-series. We demonstrate a continuous-tilting method that can record a tilt-series in seconds (about 100x faster than current methods), but with loss of details finer than ∼4 nm. We also demonstrate a fast-incremental method that can record a tilt-series about 10x faster than current methods and with similar resolution. We characterize the utility of both methods in real biological electron cryotomography workflows. We identify opportunities for further improvements in hardware and software and speculate on the impact such advances could have on structural biology.
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