title: AutoTag and AutoSnap: Standardized, semi-automatic capture of regions of interest from whole slide images.
abstract: 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 interest (ROIs) were selected by manual operations within a tumor by using a systematic uniform random sampling (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.
authors.affiliation: Laboratory of Physiopharmacology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium ; HistoGeneX NV, ZNA Middelheim Campus, Lindendreef 1, B-2020 Antwerp, Belgium.
authors.forename: Koen M
authors.initials: KM
authors.lastname: Marien
authors.affiliation: HistoGeneX NV, ZNA Middelheim Campus, Lindendreef 1, B-2020 Antwerp, Belgium.
authors.forename: Luc
authors.initials: L
authors.lastname: Andries
authors.affiliation: HistoGeneX NV, ZNA Middelheim Campus, Lindendreef 1, B-2020 Antwerp, Belgium.
authors.forename: Stefanie
authors.initials: S
authors.lastname: De Schepper
authors.affiliation: HistoGeneX NV, ZNA Middelheim Campus, Lindendreef 1, B-2020 Antwerp, Belgium.
authors.forename: Mark M
authors.initials: MM
authors.lastname: Kockx
authors.affiliation: Laboratory of Physiopharmacology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium.
authors.forename: Guido R Y
authors.initials: GR
authors.lastname: De Meyer
keywords: AutoTag and AutoSnap; Automation; Computer-assisted image processing; Microvessels; Selection bias; Stereology; Whole slide image