PURPOSE: A ground-truth validation platform was developed to provide spatial correlation between ultrasound (US), temperature measurements and histopathology images to validate US based thermal ablation monitoring methods. METHOD: The test-bed apparatus consists of a container box with integrated fiducial lines. Tissue samples are suspended within the box using agar gel as the fixation medium. Following US imaging, the gel block is sliced and pathology images are acquired. Interactive software segments the fiducials as well as structures of interest in the pathology and US images. The software reconstructs the regions in 3D space and performs analysis and comparison of the features identified from both imaging modalities. RESULTS: The apparatus and software were constructed to meet technical requirements. Tissue samples were contoured, reconstructed and registered in the common coordinate system of fiducials. There was agreement between the sample shapes, but systematic shift of several millimeters was found between histopathology and US. This indicates that during pathology slicing shear forces tend to dislocate the fiducial lines. Softer fiducial lines and harder gel material can eliminate this problem. CONCLUSION: Viability of concept was presented. Despite our straightforward approach, further experimental work is required to optimize all materials and customize software.