Biostructural Mechanism Laboratory, RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan.
Biostructural Mechanism Laboratory, RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan; Advanced Electron Microscope Development Unit, RIKEN-JEOL Collaboration Center, RIKEN Baton Zone Program, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan.
J Struct Biol. 2020 Jan 1;209(1):107414. doi: 10.1016/j.jsb.2019.107414. Epub 2019 Nov 4.
Cryo-EM technology has developed to the point of high-throughput structure determination of biological macromolecules embedded in vitreous ice. Nonetheless, challenging targets need extensive sample screening, often of many cryo-EM sample grids prepared under various conditions. We have designed and made tools for manipulating sample grids in storage cases. These tools are made of a plastic fiber using a wide-use 3D printer, a fused deposition modeling type, and polished under acetone gas. A grid case stacker organizes many frozen-hydrated cryo-EM grids and the stackers can be piled up inside a standard 50 mL centrifuge tube. We have also introduced tools that facilitate handling of grid cases under liquid nitrogen and a stocker of the grid retainers contained in a CRYO ARM electron microscope. Blueprints of the tools named CryoGridTools are available from a GitHub site.
冷冻电镜技术已经发展到可以高通量地确定嵌入玻璃态冰中的生物大分子的结构。尽管如此,具有挑战性的目标仍需要广泛的样本筛选,通常需要制备许多在不同条件下的冷冻电镜样本网格。我们设计并制造了用于在储存盒中操作样本网格的工具。这些工具由一种使用广泛的 3D 打印机、熔融沉积建模类型的塑料纤维制成,并在丙酮气体下进行抛光。网格盒堆叠器可组织多个冷冻水合的冷冻电镜网格,并且堆叠器可以堆放在标准的 50mL 离心管内。我们还引入了一些工具,方便在液氮下处理网格盒和在 CRYO ARM 电子显微镜中存放网格固定器的储存架。名为 CryoGridTools 的工具的蓝图可从 GitHub 网站获得。