Cipriani Florent, Röwer Martin, Landret Christophe, Zander Ulrich, Felisaz Franck, Márquez José Antonio
Grenoble Outstation, European Molecular Biology Laboratory, 6 Rue Jules Horowitz, 38042 Grenoble, France.
Acta Crystallogr D Biol Crystallogr. 2012 Oct;68(Pt 10):1393-9. doi: 10.1107/S0907444912031459. Epub 2012 Sep 18.
The use of automated systems for crystallization and X-ray data collection is now widespread. However, these two steps are separated by the need to transfer crystals from crystallization supports to X-ray data-collection supports, which is a difficult manual operation. Here, a new approach is proposed called CrystalDirect (CD) which enables full automation of the crystal-harvesting process. In this approach, crystals are grown on ultrathin films in a newly designed vapour-diffusion crystallization plate and are recovered by excision of the film through laser-induced photoablation. The film pieces containing crystals are then directly attached to a pin for X-ray data collection. This new method eliminates the delicate step of `crystal fishing', thereby enabling full automation of the crystal-mounting process. Additional advantages of this approach include the absence of mechanical stress and that it facilitates handling of microcrystals. The CD crystallization plates are also suitable for in situ crystal screening with minimal X-ray background. This method could enable the operational integration of highly automated crystallization and data-collection facilities, minimizing the delay between crystal identification and diffraction measurements. It can also contribute significantly to the advancement of challenging projects that require the systematic testing of large numbers of crystals.
自动化系统在结晶和X射线数据收集方面的应用如今已十分普遍。然而,这两个步骤因需要将晶体从结晶载体转移至X射线数据收集载体而被分隔开来,这是一项困难的手动操作。在此,提出了一种名为CrystalDirect(CD)的新方法,它能够实现晶体收获过程的完全自动化。在这种方法中,晶体生长在新设计的气相扩散结晶板中的超薄膜上,并通过激光诱导光烧蚀切除薄膜来回收晶体。然后,将含有晶体的薄膜碎片直接附着到用于X射线数据收集的针上。这种新方法省去了“捞晶体”这一精细步骤,从而实现了晶体安装过程的完全自动化。该方法的其他优点包括不存在机械应力,且便于处理微晶。CD结晶板也适用于具有最小X射线背景的原位晶体筛选。这种方法可以实现高度自动化的结晶和数据收集设施的操作整合,最大限度地减少晶体识别和衍射测量之间的延迟。它还可以为需要对大量晶体进行系统测试的具有挑战性的项目的推进做出重大贡献。