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

 描述（由申请人提供）：我们的实验室以开发和广泛传播流式细胞术技术而闻名，从开发第一台流式细胞仪开始，此后继续对仪器本身和软件进行创新性改进，现在取代并大大扩展了初始功能。我们早期的工作，我们的实验室是著名的，包括从硬件到软件的荧光补偿，现在无处不在的“概率”轮廓显示二维流数据的发展，和“Logicle”（又名“双指数”）显示轴的发展，是迅速变得无处不在。在这一传统的基础上，我们将注意力转向自动化荧光补偿和流式数据分析，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R 01的支持下，我们在这里寻求更新，这个项目已经产生了一系列用户友好的自动化软件“工具”，提供了可访问的，统计上适当的方法来处理和分析流量数据。这些包括1）基于统计学的自动荧光补偿; 2）基于统计学的分析软件，其降低了门控决策的主观性;以及3）用于开发和应用门控模型的直接方法，其可以在数据集内和跨数据集客观地应用。这个当前版本仍然缺乏我们建议在这里添加的重要功能。然而，我们已经开始在我们自己的实验室中使用它，并将其分发给有限的流量用户组，效果良好。按照这个轨迹，我们计划开始使这个版本很快免费提供给非营利机构（.edu，.org，.gov等）的用户，并在我们实验室增加新功能时保持发布的版本最新。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/改进几个自动化仪器和分析功能，从而显着增加其范围和易用性。整个软件包将促进流式细胞术在所有领域的工作。然而，它将为HIV研究和针对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和加速获得细胞因子和其他T细胞亚群标记物中诱导的变化的统计学有效比较，以响应病原体及其替代物的体内或离体模拟。

 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.