Accurate positioning of the naked eye is a prerequisite in many ophthalmic measurement and intervention systems. In most of these procedures the eyeball is stabilized through fixation of a target. Fixation is initiated, as well as sustained by a whole set of oculomotor processes: saccadic movements and drift, as well as vergence. Target appearance influences sustained fixation, but the influence of target information on the initiation of fixation has not been evaluated in detail. The current study evaluates the accuracy of fixation initiation as well as sustained fixation under refractive error. Twenty-one ametrope subjects repeatedly fixated a laser speckle-based fixation target for an extended duration of 3 s. Fixational area, fixational saccade rate as well as fixational saccade amplitude were analyzed during two different time intervals, namely in fixation initiation and sustained fixation. Fixation initiation was evaluated within the first 500 ms of fixation, whereas sustained fixation was evaluated 1 s after the eyes were directed toward the fixation target. During fixation initiation, fixation accuracy decreased in comparison to sustained fixation; fixational saccades occurred more frequently, and with larger amplitudes. During sustained fixation, an impact of refractive error was shown. With increasing refractive error, fixational saccade amplitude and fixational saccade rate increased. Fixational area increased in high ametropia through larger and more frequent saccades, but at a rather moderate rate of 10 arcmin/diopter (dpt) radial increase.