Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Methods Mol Biol. 2021;2215:299-307. doi: 10.1007/978-1-0716-0966-8_14.
Here, we present a strategy to identify microcrystals from initial protein crystallization screen experiments and to optimize diffraction quality of those crystals using negative stain transmission electron microscopy (TEM) as a guiding technique. The use of negative stain TEM allows visualization along the process and thus enables optimization of crystal diffraction by monitoring the lattice quality of crystallization conditions. Nanocrystals bearing perfect lattices are seeded and can be used for MicroED as well as growing larger crystals for X-ray and free electron laser (FEL) data collection.
在这里，我们提出了一种从初始蛋白质结晶筛选实验中识别微晶的策略，并使用负染色透射电子显微镜 (TEM) 作为指导技术来优化这些晶体的衍射质量。负染色 TEM 的使用允许沿过程进行可视化，从而通过监测结晶条件的晶格质量来优化晶体衍射。具有完美晶格的纳米晶体被播种，可用于 MicroED 以及生长更大的晶体以进行 X 射线和自由电子激光 (FEL) 数据收集。
