National Resource for Automated Molecular Microscopy, Simons Electron Microscopy Center, New York Structural Biology Center, New York, NY, USA.
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA.
Nat Methods. 2018 Oct;15(10):793-795. doi: 10.1038/s41592-018-0139-3. Epub 2018 Sep 24.
Most protein particles prepared in vitreous ice for single-particle cryo-electron microscopy (cryo-EM) are adsorbed to air-water or substrate-water interfaces, which can cause the particles to adopt preferred orientations. By using a rapid plunge-freezing robot and nanowire grids, we were able to reduce some of the deleterious effects of the air-water interface by decreasing the dwell time of particles in thin liquid films. We demonstrated this by using single-particle cryo-EM and cryo-electron tomography (cryo-ET) to examine hemagglutinin, insulin receptor complex, and apoferritin.
大多数在玻璃态冰中制备用于单颗粒冷冻电子显微镜（cryo-EM）的蛋白质颗粒会被吸附到气-水或基底-水界面上，这可能导致颗粒采用优先取向。通过使用快速浸入式冷冻机器人和纳米线网格，我们能够通过减少颗粒在薄液膜中的停留时间来降低气-水界面的一些有害影响。我们通过使用单颗粒冷冻电子显微镜和冷冻电子断层扫描（cryo-ET）来检查血凝素、胰岛素受体复合物和脱铁蛋白来证明这一点。
