Pachl Petr, Coudray Léna, Vincent Romain, Nilles Léa, Scheer Hélène, Ritzenthaler Christophe, Fejfarová Adéla, Řezáčová Pavlína, Engilberge Sylvain, Sauter Claude
CNRS, Architecture et Réactivité de l'ARN, UPR 9002, Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, France.
Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Praha 6, Czech Republic.
FEBS Open Bio. 2025 Apr;15(4):532-541. doi: 10.1002/2211-5463.13932. Epub 2024 Nov 21.
The production of high-quality crystals is a key step in crystallography in general, but control of crystallization conditions is even more crucial in serial crystallography, which requires sets of crystals homogeneous in size and diffraction properties. This protocol describes the implementation of a simple and user-friendly microfluidic device that allows both the production of crystals by the counter-diffusion method and their in situ analysis by serial crystallography. As an illustration, the whole procedure is used to determine the crystal structure of three proteins from data collected at room temperature at a synchrotron radiation source.
一般来说,高质量晶体的制备是晶体学中的关键步骤,但在串行晶体学中,结晶条件的控制更为关键,因为串行晶体学需要尺寸和衍射特性均一的晶体组。本方案描述了一种简单且用户友好的微流控装置的应用,该装置既能通过反扩散法制备晶体,又能通过串行晶体学对其进行原位分析。作为示例,整个过程用于根据在同步辐射源室温下收集的数据确定三种蛋白质的晶体结构。