Objective To study the clinical characteristics and differentiation of stroke mimics during super early intravenous thrombolysis of acute ischemic stroke. Methods The clinical data of patients who received intravenous thrombolysis between Sep. 2013 and Feb. 2015 in Changhai Hospital were retrospectively analyzed. And those with stroke mimics were identified and were compared with those with strokes; the clinical symptoms and laboratory findings were compared between the two groups. Results A total of 212 patients received intravenous thrombolysis, and 7 (3.3%) of them were identified as having stroke mimics. There were no notable differences in the baseline characteristics between mimics and stroke groups. Psychiatric and dementia history were of great value for differentiation of stroke mimics from strokes. MRI, vascular assessment, electroencephalogram (EEG), blood and cerebrospinal fluid assay greatly contributed to the final diagnosis of stroke mimics. Conclusion Only few patients receiving intravenous thrombolysis have been confirmed to have stroke mimics. Medical history combined with MRI may be of great value for differentiation of mimics and strokes.