Background: Post-stroke anxiety (PSA) is a common neuropsychiatric affective disorder occurring after a stroke. Animal experiments have indicated that serum S-100β levels are closely related to anxiety disorder. No clinical study has been done to explore the relationship between serum S-100β levels and anxiety symptoms in patients with acute stroke. The aim of our study was to investigate the association between serum S-100β levels and PSA. Methods: One hundred twenty-six acute stroke patients were recruited and followed up for 1 month. Blood samples were collected within 24 h after admission. The levels of serum S-100β were measured by enzyme-linked immunosorbent assays. Patients with significant clinical symptoms of anxiety and a Hamilton Anxiety Rating Scale score >7 at 1 month after stroke were diagnosed as PSA. Results: Serum S-100β levels in the non-PSA group were lower than the PSA group (838.97 (678.20-993.59) ng/L vs. 961.87 (796.09-1479.59) ng/L, Z = À2.661, P = 0.008). In multivariate analyses, we found that decreased risk of PSA was associated with low tertile serum S-100β levels (≤753.8 ng/L, OR 0.062, 95% CI 0.008-0.475, P = 0.007). Conclusions: Low serum S-100β levels at admission may be associated with the decreased risk of PSA.