It might be difficult for clinicians and scientists to identify comprehensively the major research topics given the large number of publications. A bibliometric report that identifies the most-cited articles within the body of the relevant literature may provide insight and guidance for readers toward scientific topics that are considered important for researchers and all relevant workers of academia. To our knowledge, there is a lack of an overall evaluation of the most-cited articles and hence of a comprehensive review of major research topics in neuroscience. The present study was therefore proposed to analyze and characterize the 100 most-cited articles in neuroscience. Based on data provided from Web of Science, the 100 most-cited articles relevant to neuroscience were identified and characterized. Information was extracted for each included article to assess for the publication year, journal published, impact factor, adjusted impact factor, citation count (total, normalized, and adjusted), reference list, authorship and article type. The total citation count for the 100 most-cited articles ranged from 7,326 to 2,138 (mean 3087.0) and the normalized citation count ranged from 0.163 to 0.007 (mean 0.054). The majority of the 100 articles were research articles (67%) and published from 1996 to 2000 (30%). The author and journal with the largest share of these 100 articles were Stephen M. Smith ( = 6) and ( = 13) respectively. Among the 100 most-cited articles, 37 were interlinked via citations of one another, and they could be classified into five major topics, four of which were scientific topics, namely neurological disorders, prefrontal cortex/emotion/reward, brain network, and brain mapping. The remaining topic was methodology. Interestingly 41 out of 63 of the rest, non-interlinked articles could also be categorized under the above five topics. Adjusted journal impact factor among these 100 articles did not appear to be associated with the corresponding adjusted citation count. The current study compiles a comprehensive list and analysis of the 100 most-cited articles relevant to neuroscience that enables the comprehensive identification and recognition of the most important and relevant research topics concerned.