Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, United States.
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, United States; Beckman Institute, California Institute of Technology, Pasadena, CA 91125, United States.
J Struct Biol. 2019 Feb 1;205(2):163-169. doi: 10.1016/j.jsb.2018.12.008. Epub 2019 Jan 11.
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.
利用配备单轴夹具的新型 Titan Krios 载物台,我们开发了两种加速倾斜系列采集的方法。我们展示了一种连续倾斜的方法,可以在几秒钟内记录倾斜系列,但会损失小于 ∼4nm 的细节。我们还展示了一种快速递增的方法,可以比当前方法快几倍地记录倾斜系列,并且具有相似的分辨率。我们在真实的生物电子断层扫描工作流程中对这两种方法的实用性进行了表征。我们确定了在硬件和软件方面进一步改进的机会,并推测了这些进展对结构生物学的影响。