Many studies have been carried out to assess the variability of sleep parameters. The first night effect is one of the most important factors in this variability and has been extensively studied. However, the readaptation phenomenon when subjects returned to the sleep laboratory after spending a certain period of time at home has been not systematically evaluated. To investigate this phenomenon across multiple sleep laboratory sessions, polysomnographic data from 12 healthy young subjects for 12 nights (three periods each of 4 consecutive nights, with a minimum of 1 month between them) were collected. The first night effect was present only in the first night of the first period (“very first night”) and was significant only for REM sleep-related variables. We conclude that the results from the first nights of consecutive periods within a specific protocol with healthy young subjects need not be discarded in subsequent analyses.