Westphall Michael S, Lee Kenneth W, Hemme Colin, Salome Austin Z, Mertz Keaton, Grant Timothy, Coon Joshua J
bioRxiv. 2023 Jul 22:2023.07.21.550105. doi: 10.1101/2023.07.21.550105.
We describe an apparatus for the cryogenic landing of particles from the ion beam of a mass spectrometer onto transmission electron microscope grids for cryo-electron microscopy. This system also allows for the controlled formation of thin films of amorphous ice on the grid surface. We demonstrate that as compared to room temperature landings, use of this cryogenic landing device greatly improves the structural preservation of deposited protein-protein complexes. Further, landing under cryogenic conditions can increase the diversity of particle orientations, allowing for improved 3D structural interpretation. Finally, we conclude that this approach allows for the direct coupling of mass spectrometry with cryo-electron microscopy.
我们描述了一种用于将质谱仪离子束中的粒子低温沉积到用于冷冻电子显微镜的透射电子显微镜网格上的装置。该系统还能在网格表面可控地形成非晶冰薄膜。我们证明,与室温沉积相比,使用这种低温沉积装置能极大地改善沉积的蛋白质 - 蛋白质复合物的结构保存。此外,在低温条件下沉积可增加粒子取向的多样性,从而改善三维结构解析。最后,我们得出结论,这种方法可实现质谱与冷冻电子显微镜的直接联用。