The concentration of neuron-specific enolase (NSE) in serum and cerebrospinal fluid (CSF) has been used as a biomarker in some cancers and, more recently, in neurodegenerative diseases. Pre-analytical conditions are very important for the quality of returned results. In this study, we evaluated the effects of storage conditions (temperature and duration of storage) and hemolysis on the concentration of NSE in serum and CSF. Our results demonstrate that samples for NSE measurement may be stored at -80 degrees C for no more than 6 months in the case of CSF and 9 months in the case of serum samples. Even invisible hemolysis may increase NSE levels in samples. Consequently, an index of hemolysis should be determined before deciding whether or not to perform NSE measurement.