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

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 用于晶体质量筛选和数据收集的样品的安装通常是有问题的。 传统上需要手动操作水晶才能将其正确地放置在底座内。晶体通常因物理接触和暴露于空气而损坏。 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Hauptmann Woodward研究所合作开发了与SAM系统兼容的新型样品容器capette。capette将用于低温和室温下的晶体生长和x射线衍射实验，减轻直接样品操作。 capette将用作手动移液器和自动液体处理机器人的移液器吸头，使晶体生长溶液能够轻松地吸入内部。 对于X射线数据收集，Cappette的较大端安装在金属底座上，并且组件安装在SSRL卡匣或单圆盘样品存储容器内。当用于X射线数据收集时，capette产生低X射线背景，因此可以从capette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，与一家公司开始安排生产一种在较小直径部分具有更薄壁的capette，该部分用于容纳用于数据收集的样品。 这种新的卡佩特将通过融合一个非常薄壁小直径挤出管到一个更大的直径注塑基地。 今年使用束时间来测试该公司提供的哪些薄壁管具有最佳的数据收集性能。 发现capette的直径应该是200微米或更小，以便于样品在X射线束中居中和适当的低温冷却。

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.