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首页> 外文期刊>Acta crystallographica. Section D, Structural biology. >Shake-it-off: a simple ultrasonic cryo-EM specimen-preparation device
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Shake-it-off: a simple ultrasonic cryo-EM specimen-preparation device

机译：抖掉身上的土:一个简单的超声波低温电子显微镜样品制备设备

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摘要

Although microscopes and image-analysis software for electron cryomicroscopy(cryo-EM) have improved dramatically in recent years, specimen-preparationmethods have lagged behind. Most strategies still rely on blotting microscopegrids with paper to produce a thin film of solution suitable for vitrification. Thisapproach loses more than 99.9% of the applied sample and requires severalseconds, leading to problematic air-water interface interactions for macro-molecules in the resulting thin film of solution and complicating time-resolvedstudies. Recently developed self-wicking EM grids allow the use of smallvolumes of sample, with nanowires on the grid bars removing excess solution toproduce a thin film within tens of milliseconds from sample application tofreezing. Here, a simple cryo-EM specimen-preparation device that usescomponents from an ultrasonic humidifier to transfer protein solution onto aself-wicking EM grid is presented. The device is controlled by a Raspberry Pisingle-board computer and all components are either widely available or can bemanufactured by online services, allowing the device to be constructed inlaboratories that specialize in cryo-EM rather than instrument design. Thesimple open-source design permits the straightforward customization of theinstrument for specialized experiments.
机译：尽管显微镜和图像分析软件电子cryomicroscopy(低温电子显微镜)近年来显著改善,specimen-preparationmethods落后。大多数策略仍然依靠印迹microscopegrids用纸生产薄膜适用于玻璃化的解决方案。Thisapproach失去超过99.9%的应用示例,需要severalseconds,导致有问题的空气与界面交互大分子在生成的薄膜解决方案和time-resolvedstudies复杂化。最近开发self-wicking EM电网允许的使用smallvolumes样本,纳米线在网格上酒吧去除多余的解决方案创造出一个薄膜在几十毫秒内从示例应用程序tofreezing。简单的低温电子显微镜样品制备设备usescomponents从超声波加湿器转运蛋白的解决方案到aself-wicking EM提出了网格。覆盆子Pisingle-board电脑和所有组件是广泛使用的或bemanufactured通过在线服务,允许设备构造inlaboratories专门从事低温电子显微镜而不是乐器设计。简单的定制theinstrument专业实验。

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