Quantitative assessment of HIV or SIV viral DNA in laser capture microdissected immune cells

• Calculates the fraction of captured CD4+ T cells or macrophages harboring HIV or SIV vDNA. 
• Captures up to 3,000 individual CD4+ T cells or macrophages. 
• Annotates tissue regions of interest and specific cell populations to highlight the specific anatomical tissue compartments. 
• Performs laser capture microdissection. 
• Provides relevant data electronically, including a report with interpretations and conclusions. 

The process: 

• CD4 (DAB) and CD68/CD163 (FastRed) double immunohistochemistry is performed, allowing for the specific identification of both CD4+ T cell and myeloid/macrophage populations. 
• A high-resolution digital image of each stained sample is generated using the Aperio ScanScope CS System. 
• Under the guidance of a subject matter expert, the core annotates tissue regions of interest and specific cell populations (i.e., CD4+ T cells or CD68+/CD163+ macrophages) to highlight the specific anatomical tissue compartments and specify the cell populations to be captured by laser capture microdissection. 
• Laser capture microdissection is performed on an ArcturusXT Laser Capture Microdissection System. 
• Captured CD4+ T cells or macrophages are treated using the Arcturus PicoPure DNA Extraction kit. 
• Quantitative PCR is performed for HIV or SIV vDNA (Gag) and the cellular gene CCR5 for cell equivalent enumeration. 

Includes sufficient basic consultation with a subject matter expert.