BIOMEDICAL RESEARCH SUPPORT GRANT

逆转录病毒研究中心的细胞计数实验室要求 资金用于购买锚定细胞分析和分选（ACAS）系统， 由密歇根州奥克莫斯的Meridian Instruments制造的ACAS型号570。 ACAS系统将允许研究人员分析粘附细胞， 方法类似于现在使用流式细胞仪对非粘附细胞进行的方法， 细胞仪 ACAS 570将成为该流程的配套仪器 细胞计数器（Coulter EPICS 753）目前在细胞计数中使用 实验室 这两种仪器将为研究人员提供更大的 研究细胞特性和功能的能力 in their其individual个别projects项目. ACAS 570有几个理想的 这些能力对于流式细胞仪来说是有问题的。 最 其中明显的是它能够分析活的贴壁细胞， 通过荧光可测量的参数，而无需制备单一的 细胞悬液 分离粘附细胞通常会剥离细胞膜， 被分析的蛋白质，并干扰细胞与细胞之间， 底层相互作用 ACAS 570配备了一个电机驱动 载物台、激光光学和荧光检测系统以及微计算机 其控制原位染色细胞的扫描和分析 用荧光抗体或其他荧光探针。 计算机 允许随着时间的推移重复观察相同的细胞。 激光 系统可以被编程为自动杀死预定义的细胞， 细胞亚群被分离用于未来的生长和表征。 能够描述特定变化随时间的变化， 分选贴壁细胞是ACAS的独特功能， 使用流式细胞术充分进行。 我们建议使用ACAS 570 在各种旨在表征细胞亚群的项目中， 确定细胞对刺激如病毒和病毒蛋白的反应， 激素、DNA诱变剂、药物、毒素和细胞因子。 的许多项目 需要ACAS的细胞分离和分选功能。

The Cytometry Laboratory of the Center for Retrovirus Research requests funds to purchase the Anchored Cell Analysis and Sorting (ACAS) system, the ACAS Model 570 manufactured by Meridian Instruments, Okemos, Michigan. The ACAS system will permit investigators to analyze adherent cells in a way similar to what is now performed on nonadherent cells using flow cytometry. The ACAS 570 will be a companion instrument to the flow cytometer (Coulter EPICS 753) presently in service in the Cytometry Laboratory. Together, the two instruments will give researchers greater capability to study cell characteristics and functions under investigation in their individual projects. The ACAS 570 has several desirable capabilities that are problematic for the flow cytometer. The most obvious of these is its ability to analyze live adherent cells for parameters measurable by fluorescence without the need to prepare single cell suspensions. Detaching adherent cells often strips the membranes of the very proteins to be analyzed, and disturbs cell to cell and cell to substratum interactions. The ACAS 570 is equipped with a motor-driven stage, laser optics and fluorescence detection system and micro-computer which controls scaning and analysis of fields of cells stained in situ with fluorescent antibodies or other fluorescent probes. The computer permits repeated observations of the same cells over time. The laser system can be programmed to kill predefined cells automatically allowing subsets of cells to be isolated for future growth and characterization. The ability to characterize specific changes over time and to isolate or sort adherent cells are unique features of the ACAS that cannot be performed adequately using flow cytometry. We propose to use the ACAS 570 in a variety of projects directed at characterizing cell subsets and determining cell responses to stimuli such as virus and virus protein, hormones, DNA mutagens, drugs, toxins and cytokines. Many of the projects require the cell isolation and sorting functions of the ACAS.