Novel CD4+ T Cell Specific Immunohistochemistry Detection and Analysis

• Provides comprehensive quantitative image analysis for CD4+ T cells. 
• Uses masking of non-T cell CD4 in fixed tissues from virally infected and uninfected specimens. 
• Generates a high-resolution digital image of each double immunohistochemistry stained sample. 
• Provides relevant data electronically, including a report with interpretations and conclusions. 

The process: 

• The Tissue Analysis Core processes, embeds, and performs microtomy on fixed-tissue samples presented in ethanol. 
• Slides are stained for CD4 & CD68/CD163 markers by immunohistochemistry for macrophages/dendritic cells. 
• To ensure consistency among assay batches, we have automated a major portion of the double immunohistochemistry assay using the IntelliPATH autostainer. 
• A high-resolution digital image of each double immunohistochemistry stained sample will be generated using the Aperio ScanScope CS System. 
• Regions of interest of a defined size will be highlighted on each digital image using the Aperio ImageScope software regions tool. 
• High-resolution images of regions of interest will be generated using a custom-written software program created by the Imaging and Visualization Group that automatically extracts regions of interest from the digital slide annotation layer. 
• Under the guidance of a subject matter expert, the core will provide comprehensive quantitative image analysis using a custom series of unique actions in Photoshop with Fovea tools to determine the percent area in each high-resolution image that is occupied by CD4+ T cells. 

Includes sufficient basic consultation with a subject matter expert.