对你的网格进行冷冻循环：快速冷冻玻璃化并重复使用裁剪后的网格，以推动冷冻电镜技术的普及。
CryoCycle your grids: Plunge vitrifying and reusing clipped grids to advance cryoEM democratization.
作者信息
Serbynovskyi Viacheslav, Wang Jing, Chua Eugene Yd, Ishemgulova Aygul, Alink Lambertus M, Budell William C, Johnston Jake D, Dubbeldam Charlie, Gonzalez Fabio A, Rozovsky Sharon, Eng Edward T, de Marco Alex, Noble Alex J
机构信息
Simons Electron Microscopy Center, New York Structural Biology Center, NY, NY, USA.
Department of Cellular and Molecular Physiology & Biophysics, Columbia University, New York, NY, USA.
出版信息
Res Sq. 2024 Jul 2:rs.3.rs-4415026. doi: 10.21203/rs.3.rs-4415026/v1.
CryoEM democratization is hampered by access to costly plunge-freezing supplies. We introduce methods, called CryoCycle, for reliably blotting, vitrifying, and reusing clipped cryoEM grids. We demonstrate that vitreous ice may be produced by plunging clipped grids with purified proteins into liquid ethane and that clipped grids may be reused several times for different protein samples. Furthermore, we demonstrate the vitrification of thin areas of cells prepared on gold-coated, pre-clipped grids.
低温电子显微镜技术的普及受到昂贵的骤冷设备获取的限制。我们引入了一种名为CryoCycle的方法，用于可靠地吸干、玻璃化并重复使用裁剪后的低温电子显微镜网格。我们证明，将含有纯化蛋白质的裁剪网格投入液态乙烷中可产生玻璃态冰，并且裁剪后的网格可多次重复用于不同的蛋白质样品。此外，我们还展示了在镀金的预裁剪网格上制备的细胞薄区域的玻璃化。
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