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The aim of the study was to evaluate the role of lipid layer thickness and corneal sensation in the development of blinking in neonates. The study group comprised sixty-four neonates and infants (mean age 27.5 ± 15 (sd) weeks, range 3.4–52) whose mothers were attending a general practice healthy baby clinic. Spontaneous eye-blink activity was determined from digital videographic recordings; tear film lipid layer morphology wasexamined using interference patterns produced by the Keeler Tearscope™ Plus over a five-point grading scale (higher grades are associated with thick and stable lipid films); corneal sensation threshold was assessed with the Non-Contact Corneal Aesthesiometer (NCCA), using the eye-blink response as an objective indication that the cooling stimulus had been felt; palpebral aperture dimensions were measured using calibrated digital still images of the eye in the primary position. The overall mean spontaneous blink-rate was found to be 3.6 (± 0.3) blinks min−1, and the mean interblink time was 21.6 (± 2.8) s. The lowest blink-rates were observed in the 0–17-week age group (average 2 blinks min−1). The blink-rate showed a highly significant correlation with age (r = 0.46, P < 0.01). The overall mean lipid layer grading was 3.6 (± 0.2 SE) arbitrary units. Higher grades were found in the newborn and the mean grading score reduced with age (P < 0.01). The mean sensation threshold to blink (TTB) was 0.69 (0.04 SE) mbar, which did not differ from a control group of older subjects (P > 0.05). There was a rapid increase in palpebral aperture length and width from birth to 1 year old, with surface area increasing by 50% over the same period. We concluded that the low rate of spontaneous eye blink activity in neonates is associated with a thick stable lipid layer that may be a function of a small palpebral aperture. Furthermore, neonates appear to have the capacity to detect ocular surface cooling, which is a major trigger for spontaneous blinking.