HIGH-THROUGHPUT SOLUTION SCATTERING DATA COLLECTION SYSTEM

高通量解决方案散射数据采集系统

• 批准号: 7954287
• 负责人: HIROTSUGU TSURUTA
• 金额: $ 1.86万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954287
• 关键词: Adopted; Air; Aliquot; Aluminum; Blood capillaries; Cells; Computer Retrieval of Information on Scientific Projects Database; Data Collection; Development; Ethanol; Flushing; Funding; Grant; Institution; Liquid substance; Manuals; Methods; Needles; Quartz; Radiation; Recovery; Research; Research Personnel; Resources; Risk; Sampling; Scheme; Series; Solutions; Source; Staging; Syringes; System; Teflon; Time; Translations; Tube; United States National Institutes of Health; Upper arm; Water; capillary; data acquisition; instrumentation; purge; structural biology; synchrotron radiation

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. The development of instrumentation for an automated high throughput data collection system for solution x-ray scattering continues, combining a quartz capillary cell and a motorized sample changer, which combines a refrigerated aluminum block accepting any of the standard micro-plates or several PCR tube strips (up to total 8x12 tubes), with three motorized miniature linear translation stages whose vertical arm holds a syringe needle for sample aliquot selection and delivery. A Hamilton 560 fluid dispenser replaced Hamilton 540B to accommodate a larger number of fluid inlets and outlets required to accomplish sample cell cleaning, a newly automated feature. All above instrumentation are under Blu-ICE control. This year we adopted a completely new sample delivery scheme in which a syringe needle delivers a sample aliquot directly into a capillary cell instead of sending it via Teflon tubing. The new method substantially lowers the risk of generating air bubbles in the sample aliquot. A typical data collection sequence involves 1) the syringe needle picking up a sample aliquot on PCR tube strips, 2) the needle transferring the aliquot to the capillary cell, 3) repetitive translation of sample aliquot within the quartz capillary at around 1 microliter per second to minimize radiation damage, immediately followed by the initiation of a series of scattering data acquisition, 4) optional sample recovery to a PCR tube, 5) cleaning of the capillary and syringe needle by flushing them with 50% bleach and 50% ethanol solutions, then finally with distilled water; 6) drying of the capillary and syringe needle by compressed air purging. The automated sequence eliminates manual sample change, and leads to more efficient use of beam time.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 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。 用于溶液 X 射线散射的自动化高通量数据收集系统仪器的开发仍在继续，该系统结合了石英毛细管池和电动样品更换器，该样品更换器结合了可容纳任何标准微孔板或多个 PCR 管条的冷冻铝块（最多 8x12 管），具有三个电动微型线性平移台，其垂直臂装有注射器针头，用于样品等分试样的选择和输送。 Hamilton 560 流体分配器取代了 Hamilton 540B，可容纳完成样品池清洁（一项新的自动化功能）所需的更多流体入口和出口。以上所有仪器均受 Blu-ICE 控制。今年，我们采用了一种全新的样品输送方案，其中注射器针头将样品等分试样直接输送到毛细管细胞中，而不是通过聚四氟乙烯管输送。新方法大大降低了样品等份中产生气泡的风险。典型的数据收集顺序包括 1) 注射器针头在 PCR 管条上拾取等份样品，2) 针头将等份样品转移到毛细管池，3) 在石英毛细管内以每秒 1 微升左右的速度重复平移样品等份样品为了最大限度地减少辐射损伤，随后立即开始一系列散射数据采集，4) 可选的样品回收至 PCR 管，5) 用 50% 漂白剂冲洗毛细管和注射器针头进行清洁50%乙醇溶液，最后用蒸馏水； 6)通过压缩空气吹扫来干燥毛细管和注射器针头。自动序列消除了手动样品更换，并可以更有效地利用光束时间。