CORE--IMMUNOHISTOPATHOLOGY

免疫组织病理学核心(核心C)将提供协调和有效的设施，以便对组织样本进行可重复和可预测的组织化学处理，随后将通过自动或半自动图像分析技术进行分析。斯托克斯博士和波波维奇作为首席执行官将监督一个在组织形态计量学、细胞计数、免疫组织病理学、原位杂交、凝胶分析和放射自显影方面拥有既定记录的团队。因此，这种方法可能会产生准确和可重复的结果。我们将为该设施提供监督责任，包括安排时间表，培训项目人员进行组织准备/分析，并在必要时监督成像/显微镜设施的使用。将在本提案中使用的增强型系统(蔡司Axioplan光学显微镜和MCID 4系统的微型计算机成像软件将用于提供暗场和差分干涉对比(DIC)光学功能、免疫组织化学/免疫荧光的表观荧光照明以及促进半自动细胞计数、细胞形态分析和组织区域的分数密度分析能力的软件)，其独特之处在于，它取代了以前由个别项目研究人员为此目的而使用的各种不太灵敏、繁琐的技术。该设施的技术人员还将精通成像中心建议的硬件和软件的使用，特别是在应用于免疫细胞病理学、密度测定和荧光技术时。本提案要求增加一名技术员，为项目2-4提供支助，涉及：(1)固定方法；(2)组织切片的规划和实施；(3)适当的免疫组织化学技术；(4)数据分析/管理(如体视学技术、取样方法等)。数据存储/编目和数据呈现。该核心与核心B(统计核心)之间的密切工作关系将有助于这一技术方法在数据分析和统计程序中的应用。核心B将就适当的数据库技术或数据呈现的可转换模式和/或定量体视学程序的新软件的需要提供咨询。

The Immunohistopathology Core (Core C) will provide coordinated and efficient facilities to allow reproducible and predictable histochemical processing of tissue samples that will be subsequently analyzed by automated or semi-automated image analysis techniques. Dr. Stokes and Popovich as CO-PI's will supervise a team that has an established record in dealing with the quantitative aspects of tissue morphometrics, cell counting, immunohistopathology, in situ hybridization, gel analysis and autoradiography. Accuracy and reproducible outcomes are therefore likely to result from such an approach. We will provide oversight responsibilities for the facility including scheduling, training of project personnel in histological preparation/analysis and supervise use of the imaging/microscopic facilities as necessary. The enhanced system (Zeiss Axioplan light microscope together with microcomputer imaging software of an MCID 4 system will capacities for darkfield and differential interference contrast (DIC) optics, epi-fluorescence illumination for immunohistochemistry/immunofluorescence and software that facilitates semi-automated cell counts, cell morphometric analysis, and the ability to do fractional densitometric analysis of tissue areas) that will be used in this proposal is unique in that it replaces a variety of less sensitive, cumbersome technologies that have previously been used for this purpose by individual project investigators. Technical personnel in the facility will also be well versed in the employment of the hardware and software proposed in the imaging center, particularly as it applies to immunocytopathology, densitometry, and fluorescent techniques. An additional technician requested in the present proposal will provide support to projects 2-4 involving: (1) methods of fixation, (2) planning and execution of tissue sectioning, (3) appropriate immunohistochemical techniques, and (4) data analysis/management (e.g., stereological techniques, sampling methods, etc.) data storage/cataloging, and data presentation. This technical approach will be aided in data analysis and statistical procedures by a close working relationship between this Core and Core B (Statistical Core). Core B will advise on appropriate database technology or need for convertible modes of data presentation and/or new software for quantitative stereological procedures.