The Practical Implications of Re-Referencing in ERP Studies: The Case of N400 in the Picture–Word Verification Task

Background: The selection of an optimal referencing method in event-related potential (ERP) research has been a long-standing debate, as it can significantly influence results and lead to data misinterpretation. Such misinterpretation can produce flawed scientific conclusions, like the inaccurate localization of neural processes, and in practical applications, such as using ERPs as biomarkers in medicine, it may result in incorrect diagnoses or ineffective treatments. In line with the development and advancement of good scientific practice (GSP) in ERP research, this study sought to address several questions regarding the most suitable digital reference for investigating the N400 ERP component. Methods: The study was conducted on 17 neurotypical participants. Based on previous research, the references evaluated included the common average reference (AVE), mean earlobe reference (EARS), left mastoid reference (L), mean mastoids reference (MM), neutral infinity reference (REST), and vertex reference (VERT). Results: The results showed that all digital references, except for VERT, successfully elicited the centroparietal N400 effect in the picture–word verification task. The AVE referencing method showed the most optimal set of metrics in terms of effect size and localization, although it also produced the smallest difference waves. The most similar topographic dynamics in the N400 window were observed between the AVE and REST referencing methods. Conclusions: As the most optimal regions of interest (ROI) for the picture–word elicited N400 effect, nine electrode sites spanning from superior frontocentral to parietal regions were identified, showing consistent effects across all referencing methods except VERT.