The definitive diagnosis of breast cancer still requires obtaining a tissue sample, most commonly with an image-guided needle biopsy. High-resolution breast-dedicated positron emission mammography (PEM) has been used to guide biopsy due to its high sensitivity, specificity, and positive predictive value in detecting lesions. Although few promising methods have been proposed, these approaches require iterative procedures to verify three-dimensional (3D) needle positioning and anatomical context are not yet available. To address these challenges, we previously developed a mechatronic guidance system integrating an ultrasound (US)-guided core-needle biopsy (CNB) intervention to operate with a high-resolution PEM system. This paper presents the first assessment of system performance and phantom validation for PEM-US-guided biopsy under high-resolution PEM localization. We optimized the user interface by implementing a dynamic visualization module for 3D guidance within a simulated PEM image. Breast phantoms with targets in known 3D positions were fabricated, simulating PEM localized breast lesions and corresponding to virtual targets in the PEM image. The mechatronic guidance system was registered to the simulated PEM detector plate and real-time targeting was performed in three spatial dimensions. Biopsy procedures (N=8) were 100% successful in targets 6 mm diameter and height with a mean 3D positioning error <1.0 mm, and 0.72 mm in-plane and 0.34 mm cross-plane component errors, relative to the PEM image coordinate space. This first proof-of-concept study demonstrates a highly accurate system and feasible workflow toward PEM-USguided biopsy under high-resolution PEM localization
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