Context:The quality of images produced by whole slide imaging (WSI) scanners has a direct influence on the readers’ performance and reliability of the clinical diagnosis. Therefore, WSI scanners should produce not only high quality but also consistent quality images.Aim:We aim to evaluate reproducibility of WSI scanners based on the quality of images produced over time and among multiple scanners. The evaluation is independent of content or context of test specimen.Methods:The ultimate judge of image quality is a pathologist, however, subjective evaluations are heavily influenced by the complexity of a case and subtle variations introduced by a scanner can be easily overlooked. Therefore, we employed a quantitative image quality assessment method based on clinically relevant parameters, such as sharpness and brightness, acquired in a survey of pathologists. The acceptable level of quality per parameter was determined in a subjective study. The evaluation of scanner reproducibility was conducted with Philips Ultra-Fast Scanners. A set of 36 HercepTest™ slides were used in three sub-studies addressing variations due to systems and time, producing 8640 test images for evaluation.Results:The results showed that the majority of images in all the sub-studies are within the acceptable quality level; however, some scanners produce higher quality images more often than others. The results are independent of case types, and they match our perception of quality.Conclusion:The quantitative image quality assessment method was successfully applied in the HercepTest™ slides to evaluate WSI scanner reproducibility. The proposed method is generic and applicable to any other types of slide stains and scanners.