STOPPED FLOW TIME RESOLVED SAXS DEVELOPMENT

停止流动时间解决了 SAXS 的开发

• 批准号: 7722778
• 负责人: LIANG GUO
• 金额: $ 2.32万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722778
• 关键词: Advertising; Computer Retrieval of Information on Scientific Projects Database; Consumption; Data Quality; Data Set; Development; England; Event; Funding; Grant; Institution; KLK3 gene; Logic; Proteins; RNA; Rate; Reaction; Reagent; Research; Research Personnel; Resolution; Resources; Running; Sampling; Source; Standards of Weights and Measures; Techniques; Testing; Time; United States National Institutes of Health; cytochrome c; data acquisition; detector; instrument; posters; research study; symposium

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. Time resolved SAXS is one of the core projects in the BioCAT proposal. Development stopped flow time-resolved SAXS experiments used standard protein (cytochrome c) and RNA samples. The results demonstrate that BioCAT is capable of performing time-resolved SAXS experiments by using the SFM-400 stopped-flow instrument (Bio-Logic) combined with the Aviex CCD in streak camera mode at a time resolution of 4 ms with a reaction dead time of 1.8 ms. For each single mixing event of a sample with a reagent, 16 continuous streak frames can be recorded on the Aviex CCD chip in a single data acquisition, enabling data acquisition over a total time window of 64 ms. Longer time windows may be achieved by using larger exposure times per streak. Sample consumption can be maintained at a low level of ~ 40 ul per mixing event. RNA and protein samples at concentrations as low as ~ 1mg/ml and 5 mg/ml respectively can yield high quality data by averaging the results of 20 data sets. The results of the experiments have been presented in a poster to advertise this technique in the BSR2007 conference in England. In run 2007-3 we tested a Pilatus 100K detector borrowed from the APS detector pool in comparison to the Aviex. The data quality obtained with the Pilatus detector is considerably superior to that obtained with the Aviex detector for framing rates of 200 Hz or slower.

这个子项目是许多研究子项目中的一个 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来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 时间分辨SAXS是BioCAT提案中的核心项目之一。 开发停止流动时间分辨SAXS实验使用标准蛋白质（细胞色素c）和RNA样品。 结果表明，BioCAT能够使用SFM-400停流仪进行时间分辨SAXS实验（Bio-Logic）与Aviex CCD结合，在条纹相机模式下，时间分辨率为4 ms，反应死区时间为1.8 ms。对于样品与试剂的每个单一混合事件，在一次数据采集中，Aviex CCD芯片上可以记录16个连续的条纹帧，从而在64 ms的总时间窗口内实现数据采集。 样品消耗量可以保持在每个混合事件约40 ul的低水平。 RNA和蛋白质样品的浓度分别低至~ 1 mg/ml和5 mg/ml，通过对20个数据集的结果进行平均，可以得到高质量的数据。实验的结果已经在英国的BSR 2007会议上以海报的形式展示。在运行2007-3中，我们测试了从APS探测器池借来的Pilatus 100 K探测器，并与Aviex进行了比较。对于200 Hz或更低的成帧速率，使用Pilatus检测器获得的数据质量明显优于使用Aviex检测器获得的数据质量上级。