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Objectives: The Pupillographic Sleepiness Test (PST) measures the level of alertness based on spontaneous oscillations in pupillary size. Reference data are available for male and female adults within the age range 20-60 years. The aim of the present multicenter study was to extend the age range for reference data from 20 to 79 years., Methods: A total of 239 healthy subjects, selected from three different centers (Berlin, Regensburg, and Vienna), were administered the PST. Data were analyzed with respect to possible effects of age, gender, and time of day on the pupillary unrest index (PUI) and the pupil diameter (PD)., Results: Reliable data were available in 85.8% of the entire study sample and in 82.4% of elderly subjects (60+ years). Age-related changes were identified for the PUI and PD. There was no effect of gender on pupillometric variables as revealed by univariate analysis. The PD was found to be smaller in the afternoon compared with the morning. However, if subjects aged 60+ years were excluded from the analyses, the age-related changes disappeared and a time-of-day effect regarding the PUI became apparent., Conclusions: PUI and PD were found to decrease with increasing age. In addition, the present data show that nonreliable PST data occur more frequently in the elderly probably because of dry eyes, inadequate testing conditions, and technical resolution limitations. Thus, the PST results obtained here from elderly subjects are limited and have to be interpreted with caution. Additional research on elderly specific assessment tools is needed., (Copyright © 2012 Wiley Periodicals, Inc.)