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TitleShake-it-off: a simple ultrasonic cryo-EM specimen-preparation device.
Journal, issue, pagesActa Crystallogr D Struct Biol, Vol. 75, Issue Pt 12, Page 1063-1070, Year 2019
Publish dateDec 1, 2019
AuthorsJohn L Rubinstein / Hui Guo / Zev A Ripstein / Ali Haydaroglu / Aaron Au / Christopher M Yip / Justin M Di Trani / Samir Benlekbir / Timothy Kwok /
PubMed AbstractAlthough microscopes and image-analysis software for electron cryomicroscopy (cryo-EM) have improved dramatically in recent years, specimen-preparation methods have lagged behind. Most strategies ...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 the 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, a simple cryo-EM specimen-preparation device that uses components from an ultrasonic humidifier to transfer protein solution onto a self-wicking EM grid is presented. 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 the straightforward customization of the instrument for specialized experiments.
External linksActa Crystallogr D Struct Biol / PubMed:31793900 / PubMed Central
MethodsEM (single particle)
Resolution2.6 Å
Structure data

EMDB-20837:
Equine spleen apoferritin prepared with the Shake-it-off device
Method: EM (single particle) / Resolution: 2.6 Å

Source
  • Equus caballus (horse)

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