使用聚焦离子束研磨制备用于微晶电子衍射（MicroED）的晶体薄片的协议。
Protocol for the use of focused ion-beam milling to prepare crystalline lamellae for microcrystal electron diffraction (MicroED).
作者信息
Martynowycz Michael W, Gonen Tamir
机构信息
Howard Hughes Medical Institute, University of California Los Angeles, Los Angeles, CA 90095, USA.
Department of Biological Chemistry, University of California Los Angeles, Los Angeles, CA 90095, USA.
出版信息
STAR Protoc. 2021 Jul 28;2(3):100686. doi: 10.1016/j.xpro.2021.100686. eCollection 2021 Sep 17.
We present an in-depth protocol to reproducibly prepare crystalline lamellae from protein crystals for subsequent microcrystal electron diffraction (MicroED) experiments. This protocol covers typical soluble proteins and membrane proteins embedded in dense media. Following these steps will allow the user to prepare crystalline lamellae for protein structure determination by MicroED. For complete details on the use and execution of this protocol, please refer to Martynowycz et al. (2019a, 2020a).
我们提出了一种深入的方案，用于从蛋白质晶体中可重复地制备晶体薄片，以用于后续的微晶电子衍射（MicroED）实验。该方案涵盖了典型的可溶性蛋白质以及嵌入致密介质中的膜蛋白。按照这些步骤，用户将能够制备用于通过MicroED确定蛋白质结构的晶体薄片。有关此方案的使用和执行的完整详细信息，请参考Martynowycz等人（2019a，2020a）。
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