AutoTag and AutoSnap: Standardized, semi-automatic capture of regions of interest from whole slide images

Tumor angiogenesis is measured by counting microvessels in tissue sections at high power magnification as a potential prognostic or predictive biomarker. Until now, regions of interest1 (ROIs) were selected by manual operations within a tumor by using a systematic uniform random sampling2 (SURS) approach. Although SURS is the most reliable sampling method, it implies a high workload. However, SURS can be semi-automated and in this way contribute to the development of a validated quantification method for microvessel counting in the clinical setting. Here, we report a method to use semi-automated SURS for microvessel counting:
• Whole slide imaging with Pannoramic SCAN (3DHISTECH)
• Computer-assisted sampling in Pannoramic Viewer (3DHISTECH) extended by two self-written AutoHotkey applications (AutoTag and AutoSnap)
• The use of digital grids in Photoshop® and Bridge® (Adobe Systems)This rapid procedure allows traceability essential for high throughput protein analysis of immunohistochemically stained tissue.