PLASTIC PIPETTE TIPS AS SAMPLE MOUNTS FOR PROTEIN CRYSTALLOGRAPHY X-RAY DIFFRACT

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 用于晶体质量筛选和数据收集的样品安装通常会出现问题。传统上，需要手动操作水晶才能将其正确地放置在支架内。晶体通常会因身体接触和暴露在空气中而受损。为了简化高通量结晶设施和自动化X射线衍射数据收集，SSRL与豪普特曼·伍德沃德研究所合作开发了与SAM系统兼容的新样品容器Capette。该采样器将用于低温和室温下的晶体生长和X射线衍射实验，减少了直接的样品操作。吸头将被用作吸管尖端，带有手动移液器和自动液体处理机器人，使晶体生长的解决方案能够轻松地抽出内部。对于X射线数据采集，盖子的较大端安装在金属底座上，组件安装在SSRL盒或单冰球样品存储容器内。当用于X射线数据采集时，采集器产生低的X射线背景，因此可以从采集器内的样品中收集完整的数据集。根据前几年的材料测试和选择，与一家公司启动了安排，以生产一种在较小直径部分具有更薄壁面的盖子，该盖子容纳用于数据收集的样品。这种新的管帽将通过将一个非常薄的壁小直径挤压管熔合到一个更大直径的注塑基座上来生产。今年利用射束时间测试了该公司供应的哪种薄壁油管的数据收集性能最好。研究发现，卡普塔的直径应小于或等于200微米，以便于样品在X射线束中居中和适当的冷冻冷却。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Mounting samples for crystal quality screening and data collection can often be problematic. Manual manipulation of the crystal is traditionally required to place it properly inside a mount. Crystals are commonly damaged both by physical contact and exposure to air. To streamline high-throughput crystallization facilities and automated x-ray diffraction data collection, the capette, a new sample container compatible with the SAM system was developed at SSRL in collaboration with the Hauptmann Woodward Institute. The capette will be used for both crystal growth and x-ray diffraction experiments at cryogenic and room temperature, mitigating direct sample manipulation. The capette will be used as a pipette tip with both manual pipetters and automated liquid handling robots enabling solutions for crystal growth to be easily drawn inside. For x-ray data collection, the larger end of the cappette fits onto a metal base and the assembly fits inside the SSRL cassette or uni-puck sample storage container. When used for x-ray data collection, the capette produces a low x-ray background so complete data sets may be collected from samples within the capette. Based on materials testing and selection from previous years, arrangements were initiated with a company to produce a capette with a much thinner wall at the smaller diameter portion which holds the samples used for data collection. This new capette will be produced by fusing a very thin walled small diameter extruded tube to a larger diameter injection molded base. Beam time was used this year to test which of the thin walled tubing supplied by this company had the best properties for data collection. It was found that capettes should be 200 microns in diameter or smaller to facilitate sample centering in the x-ray beam and proper cryo-cooling.