- EMDB-20837: Equine spleen apoferritin prepared with the Shake-it-off device -
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Basic information
Entry
Database: EMDB / ID: EMD-20837
Title
Equine spleen apoferritin prepared with the Shake-it-off device
Map data
apoferritin prepared by the shake-it-off device
Sample
Complex: apoferritin
Function / homology
Function and homology information
: / intracellular sequestering of iron ion / ferric iron binding / ferrous iron binding / iron ion transport / iron ion binding / cytoplasm Similarity search - Function
Natural Sciences and Engineering Research Council (Canada)
Discovery Grant
Canada
Citation
Journal: Acta Crystallogr D Struct Biol / Year: 2019 Title: Shake-it-off: a simple ultrasonic cryo-EM specimen-preparation device. Authors: John L Rubinstein / Hui Guo / Zev A Ripstein / Ali Haydaroglu / Aaron Au / Christopher M Yip / Justin M Di Trani / Samir Benlekbir / Timothy Kwok / Abstract: 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 ...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.
History
Deposition
Oct 16, 2019
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Header (metadata) release
Dec 11, 2019
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Map release
Dec 11, 2019
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Update
Dec 2, 2020
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Current status
Dec 2, 2020
Processing site: RCSB / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: COUNTING / Digitization - Sampling interval: 14.0 µm / Number grids imaged: 1 / Number real images: 324 / Average exposure time: 30.0 sec. / Average electron dose: 21.0 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron optics
Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal magnification: 75000
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