Pathology specimen labeling errors occur for a variety of reasons. We investigated the use of barcode technology as a method to improve the accuracy of pathology specimen labeling and patient safety. We also assessed nurses’ perceptions of system quality, information quality, service quality, user satisfaction, and net benefits. Sixty-eight perioperative nurses who work in a teaching hospital in Taiwan completed the survey. Nurses scored net benefits as highly contributing to their satisfaction, whereas system quality contributed most to dissatisfaction. Further, we analyzed pathology specimen records before and after implementing the barcode system and found that specimen management errors significantly decreased. The use of a reliable barcode system could improve specimen labeling accuracy and enhance nurses’ satisfaction with this technology.