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  • Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation

    Author(s)
    Ashtiani, Dariush
    Venugopal, Hari
    Belousoff, Matthew
    Spicer, Bradley
    Mak, Johnson
    Neild, Adrian
    de Marco, Alex
    Griffith University Author(s)
    Mak, Johnson
    Year published
    2018
    Metadata
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    Abstract
    Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge ...
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    Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6–8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves.
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    Journal Title
    Journal of Structural Biology
    Volume
    203
    Issue
    2
    DOI
    https://doi.org/10.1016/j.jsb.2018.03.012
    Subject
    Biochemistry and Cell Biology not elsewhere classified
    Biochemistry and Cell Biology
    Zoology
    Publication URI
    http://hdl.handle.net/10072/380638
    Collection
    • Journal articles

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