- EMDB-10533: Horse spleen apoferritin deposited with a microfluidic sprayer -
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基本情報
登録情報
データベース: EMDB / ID: EMD-10533
タイトル
Horse spleen apoferritin deposited with a microfluidic sprayer
マップデータ
Final sharpened map
試料
複合体: apoferritin from equine spleen
機能・相同性
機能・相同性情報
: / intracellular sequestering of iron ion / ferric iron binding / ferrous iron binding / iron ion transport / iron ion binding / cytoplasm 類似検索 - 分子機能
Biotechnology and Biological Sciences Research Council
BB/P026397/1
英国
引用
ジャーナル: Acta Crystallogr D Struct Biol / 年: 2020 タイトル: Sample deposition onto cryo-EM grids: from sprays to jets and back. 著者: David P Klebl / Diana C F Monteiro / Dimitrios Kontziampasis / Florian Kopf / Frank Sobott / Howard D White / Martin Trebbin / Stephen P Muench / 要旨: Despite the great strides made in the field of single-particle cryogenic electron microscopy (cryo-EM) in microscope design, direct electron detectors and new processing suites, the area of sample ...Despite the great strides made in the field of single-particle cryogenic electron microscopy (cryo-EM) in microscope design, direct electron detectors and new processing suites, the area of sample preparation is still far from ideal. Traditionally, sample preparation involves blotting, which has been used to achieve high resolution, particularly for well behaved samples such as apoferritin. However, this approach is flawed since the blotting process can have adverse effects on some proteins and protein complexes, and the long blot time increases exposure to the damaging air-water interface. To overcome these problems, new blotless approaches have been designed for the direct deposition of the sample on the grid. Here, different methods of producing droplets for sample deposition are compared. Using gas dynamic virtual nozzles, small and high-velocity droplets were deposited on cryo-EM grids, which spread sufficiently for high-resolution cryo-EM imaging. For those wishing to pursue a similar approach, an overview is given of the current use of spray technology for cryo-EM grid preparation and areas for enhancement are pointed out. It is further shown how the broad aspects of sprayer design and operation conditions can be utilized to improve grid quality reproducibly.