Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Jahnstrasse 29, 69120 Heidelberg, Germany.
Japan Synchrotron Radiation Research Institute (JASRI), 1-1-1 Kouto, Sayo, Hyogo 679-5198, Japan.
Acta Crystallogr D Struct Biol. 2018 Oct 1;74(Pt 10):1000-1007. doi: 10.1107/S2059798318011634. Epub 2018 Oct 2.
Crystallography chips are fixed-target supports consisting of a film (for example Kapton) or wafer (for example silicon) that is processed using semiconductor-microfabrication techniques to yield an array of wells or through-holes in which single microcrystals can be lodged for raster-scan probing. Although relatively expensive to fabricate, chips offer an efficient means of high-throughput sample presentation for serial diffraction data collection at synchrotron or X-ray free-electron laser (XFEL) sources. Truly efficient loading of a chip (one microcrystal per well and no wastage during loading) is nonetheless challenging. The wells or holes must match the microcrystal size of interest, requiring that a large stock of chips be maintained. Raster scanning requires special mechanical drives to step the chip rapidly and with micrometre precision from well to well. Here, a `chip-less' adaptation is described that essentially eliminates the challenges of loading and precision scanning, albeit with increased, yet still relatively frugal, sample usage. The device consists simply of two sheets of Mylar with the crystal solution sandwiched between them. This sheet-on-sheet (SOS) sandwich structure has been employed for serial femtosecond crystallography data collection with micrometre-sized crystals at an XFEL. The approach is also well suited to time-resolved pump-probe experiments, in particular for long time delays. The SOS sandwich enables measurements under XFEL beam conditions that would damage conventional chips, as documented here. The SOS sheets hermetically seal the sample, avoiding desiccation of the sample provided that the X-ray beam does not puncture the sheets. This is the case with a synchrotron beam but not with an XFEL beam. In the latter case, desiccation, setting radially outwards from each punched hole, sets lower limits on the speed and line spacing of the raster scan. It is shown that these constraints are easily accommodated.
结晶学芯片是固定靶支撑物,由经过半导体微加工技术处理的薄膜(例如 Kapton)或晶圆(例如硅)制成,以产生可以容纳单个微晶体的井或通孔阵列,用于光栅扫描探测。虽然制造起来相对昂贵,但芯片提供了一种高效的高通量样品呈现方式,可用于同步辐射或 X 射线自由电子激光(XFEL)源的串行衍射数据收集。然而,要实现芯片的真正高效加载(每个孔一个微晶体,并且在加载过程中没有浪费)仍然具有挑战性。孔或孔必须与感兴趣的微晶体尺寸匹配,这需要大量的芯片库存。光栅扫描需要特殊的机械驱动器来快速、以微米精度从一个孔移动到另一个孔。这里描述了一种无芯片的适应方法,它基本上消除了加载和精密扫描的挑战,尽管增加了但仍然相对节俭的样品使用量。该设备仅由两片聚酯薄膜组成,晶体溶液夹在它们之间。这种片上片(SOS)三明治结构已用于在 XFEL 上进行微米大小晶体的串行飞秒结晶学数据收集。该方法也非常适合时间分辨泵浦探测实验,特别是对于长的时间延迟。SOS 三明治可以在 XFEL 光束条件下进行测量,这些条件会损坏传统的芯片,如这里所记录的那样。SOS 片可以密封样品,避免样品干燥,只要 X 射线束不刺穿片材。对于同步辐射束来说是这样的,但对于 XFEL 束来说不是。在后一种情况下,从每个打孔孔向外辐射的干燥会对光栅扫描的速度和线间距设置下限。结果表明,这些限制很容易适应。