Automated comparison of flow data from HIV and vaccine infected subjects.

 描述(申请人提供)：我们的实验室以开发和广泛传播流式细胞仪技术而闻名，从开发第一台FACS开始，然后继续对仪器本身和软件进行创新改进，现在取代并极大地扩展了最初的功能。我们实验室以其早期的工作而闻名，包括从硬件到基于软件的荧光补偿，开发现在无处不在的用于显示二维流动数据的“概率”等高线，以及开发正在迅速变得无处不在的“逻辑”(又名“双指数”)显示轴。延续这一传统，我们将注意力转向自动化荧光补偿和流动数据分析，以使各级研究人员和临床医生能够充分利用当今高维(Hi-D)流式细胞仪提供的功能。在我们寻求更新的R01的支持下，该项目已经生成了一系列用户友好的自动化软件“工具”，这些工具提供了可访问的、统计上合适的方法来处理和分析流动数据。这些包括1)基于统计的自动荧光补偿；2)基于统计的分析软件，它减少了选通决策的主观性；以及3)开发和应用可在数据集内和跨数据集客观应用的选通模型的简单方法。当前版本仍然缺乏我们建议在此处添加的重要功能。然而，我们已经开始在我们自己的实验室使用它，并将其分发给有限的流量用户群，效果良好。按照这一轨迹，我们将很快开始向非营利性机构(.edu、.org、.gov等)的用户免费提供该版本，并在我们的实验室添加新功能时保持发布版本的最新状态。我们在此请求续签资金，以使我们能够完成这项当前技术的各个方面，并添加/改进几个自动化仪器和分析功能，这些功能将显著增加其范围和易用性。整个套装将促进流式细胞仪在所有领域的工作。然而，它将为艾滋病毒研究和针对艾滋病毒和其他疾病的疫苗试验提供特别的好处，因为其强大的自动化实用程序将简化和加快对细胞因子和其他T细胞亚群标志物在响应病原体及其代理人的体内或体外模拟而诱导的细胞因子和其他T细胞亚群标志物变化的统计有效比较的访问。

 DESCRIPTION (provided by applicant): Our laboratory is well known for having developed and broadly disseminated flow cytometry technology, starting with development of the first FACS and continuing thereafter to add innovative improvements both to the instrument itself and the software that now replaces and greatly extends the initial capabilities. Our earlier work, for which our laboratory is renowned, includes the shift from hardware to software based fluorescence compensation, the development of the now-ubiquitous "probability" contours for displaying two-dimensional flow data, and the development of "Logicle" (aka "Bi-exponential") display axes that are fast becoming ubiquitous. Continuing in this tradition, we have turned our attention to automating fluorescence compensation and flow data analysis to enable researchers and clinicians at all levels to take full advantage of the capabilities offered by today's High-Dimensional (Hi-D) flow cytometry instruments. Supported by the R01 for which we seek renewal here, this project has already generated a spate of user-friendly automated software "tools" that provide accessible, statistically appropriate methods for processing and analyzing flow data. These include 1) statistically-based automated fluorescence compensation; 2) statistically- based analysis software that decreases the subjectivity of gating decisions; and 3) straightforward methods for developing and applying gating models that can be objectively applied within and across data sets. This current version still lacks important capabilities that w propose to add here. However, we have already started using it in our own laboratory and have distributed it to a limited group of flow users with good results. Following this trajectory, we pln to begin making this version available very soon at no charge to users at non-profit institutions (.edu, .org, .gov, etc.), and to keep the released version up-to-date as we add new capabilities in our laboratory. We request renewal funding here to enable us to complete aspects of this current technology and to add/improve several automated instrumentation and analysis functionalities that will significantly increase its scope and ease of use. The overall package wil facilitate flow cytometry work in all realms. However, it will offer particular benefits for HIV studies and for vaccine trials targeted at HIV and other diseases, since its robust automated utilities will simplify and speed access to statistically valid comparisons of changes induced in cytokine and other T cell subset markers in response to in vivo or ex vivo simulation by pathogens and their surrogates.