The aim of this study was to establish the relationship between OST, tear film stability as assessed by NIBUT and subjective evaluation of the lipid layer thickness in a young, asymptomatic, sample group (N=29). Non-invasive tear break-up time (NIBUT) and tear lipid layer structure were evaluated through a slit-lamp mounted Tearscope Plus. A self-calibrating infrared thermography camera was used to record two OST values (one immediately post-blink and one immediately pre the subsequent blink). The most common lipid layer pattern observed was the amorphous pattern (48.3%). Differences between post- and pre-blink OST values were observed (paired t-test; p<0.001). Significant differences of pre-blink OST values were observed between the closed marmoreal group with that from the amorphous and flow groups (Tukey post hoc test, p<0.05). There were no differences of NIBUT values between each lipid layer thickness (Kruskal-Wallis test; p=0.152). A no significant tendency for higher OST in eyes with increased NIBUT was observed. This study suggests that higher OST values could be associated with thicker tear lipid layer in normal subjects. The lack of significant results in relation with tear film stability may be due to only normal subjects were included.