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Shake-it-off: A simple ultrasonic cryo-EM specimen preparation device

By John L. Rubinstein, Hui Guo, Zev A. Ripstein, Ali Haydaroglu, Aaron Au, Christopher M. Yip, Justin M. Di Trani, Samir Benlekbir, Timothy Kwok

Posted 09 May 2019
bioRxiv DOI: 10.1101/632125 (published DOI: 10.1107/S2059798319014372)

Although microscopes and image analysis software for electron cryomicroscopy (cryo-EM) have improved dramatically in recent years, specimen preparation methods have lagged behind. Most strategies still rely on blotting microscope grids with paper to produce a thin film of solution suitable for vitrification. This approach loses more than 99.9% of the applied sample and requires several seconds, leading to problematic air-water interface interactions for macromolecules in the resulting thin film of solution and complicating time-resolved studies. Recently developed self-wicking EM grids allow use of small volumes of sample, with nanowires on the grid bars removing excess solution to produce a thin film within tens of milliseconds from sample application to freezing. Here we present a simple cryo-EM specimen preparation device that uses components from an ultrasonic humidifier to transfer protein solution onto a self-wicking EM grid. The device is controlled by a Raspberry Pi single board computer and all components are either widely available or can be manufactured by online services, allowing the device to be constructed in laboratories that specialize in cryo-EM, rather than instrument design. The simple open-source design permits straightforward customization of the instrument for specialized experiments.

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