There are two major measures of eye irritation. One is blink frequency which can be observed by human behavior. The other measures are break up time, tear flow, hyperemia (redness, swelling), tear fluid cytology, and epithelial damage (vital stains) etc., which are human beings' physiological reactions. Blink frequency is defined as the number of blinks per minute and it is associated with eye irritation. Blink frequencies are individual with mean frequencies of < 2–3 to 20–30 blinks/minute, and they depend on environmental factors including the use of contact lenses. Dehydration, mental activities, work conditions, room temperature, relative humidity, and illumination all influence blink frequency. Break-up time (BUT) is another major measure of eye irritation and tear film stability. It is defined as the time interval (in seconds) between blinking and rupture. BUT is considered to reflect the stability of the tear film as well. In normal persons, the break-up time exceeds the interval between blinks, and, therefore, the tear film is maintained. Studies have shown that blink frequency is correlated negatively with break-up time. This phenomenon indicates that perceived eye irritation is associated with an increase in blink frequency since the cornea and conjunctiva both have sensitive nerve endings that belong to the first trigeminal branch. Other evaluating methods, such as hyperemia, cytology etc. have increasingly been used to assess eye irritation.