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

 描述（由申请人提供）：我们的实验室因开发并广泛传播流式细胞术技术而闻名，从开发第一台 FACS 开始，此后继续对仪器本身和软件进行创新改进，现已取代并大大扩展了初始功能。我们的实验室因早期工作而闻名，包括从基于硬件的荧光补偿转向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数据的“概率”轮廓，以及开发迅速普及的“Logicle”（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式数据分析，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) 流式细胞术仪器提供的功能。在我们寻求更新的 R01 的支持下，该项目已经生成了大量用户友好的自动化软件“工具”，为处理和分析流量数据提供了可访问的、统计上适当的方法。其中包括 1) 基于统计的自动荧光补偿； 2) 基于统计的分析软件，减少门控决策的主观性； 3）开发和应用门控模型的简单方法，可以客观地在数据集内和数据集之间应用。当前版本仍然缺乏我们建议在此处添加的重要功能。不过，我们已经开始在自己的实验室使用它，并将其分发给有限的流量用户群体，取得了良好的效果。遵循这一轨迹，我们计划很快开始向非营利机构（.edu、.org、.gov 等）的用户免费提供此版本，并在我们实验室添加新功能时保持已发布版本的最新状态。我们在此请求更新资金，以使我们能够完成当前技术的各个方面，并添加/改进一些自动化仪器和分析功能，这将显着扩大其范围和易用性。整个软件包将促进所有领域的流式细胞术工作。然而，它将为 HIV 研究以及针对 HIV 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并加速对细胞因子和其他 T 细胞子集标记物响应于病原体及其替代物的体内或离体模拟而诱导的变化进行统计上有效的比较。

 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.