Rapid tilt-series acquisition for electron cryotomography

Rapid tilt-series acquisition for electron cryotomography
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DOI:
10.1016/j.jsb.2018.12.008
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发表时间:
2019-02-01
影响因子:
3
通讯作者:
Jensen, Grant J.
Jensen, Grant J.
中科院分区:
生物学3区
文献类型:
--
作者:
Chreifi, Georges;Chen, Songye;Jensen, Grant J.

文献摘要

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使用一个新的泰坦Krios阶段配备了一个单轴保持器,我们开发了两种方法来加速倾斜系列的收集。我们展示了一个连续倾斜的方法,可以记录在几秒钟内的倾斜系列,但细节损失比4纳米。我们还展示了一个快速增量的方法,可以记录一个倾斜系列几倍的速度比目前的方法和类似的分辨率。我们表征了这两种方法在真实的生物电子冷冻断层扫描工作流程中的实用性。我们确定了进一步改进硬件和软件的机会,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, but with loss of details finer than 4 nm. We also demonstrate a fast-incremental method that can record a tilt-series several-fold 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.