Animal allergens constitute a serious health risk in laboratory animal facilities. To assess possibilities for allergen reduction by technical and organizational measures, we studied personnel exposure to mouse urinary aeroallergens in an animal facility with a holding capacity of 30,000 cages. Short-term (2 h) and intermediate-term (12 h) stationary samples (n = 107) and short-term (2 h) personnel samples (n = 119) were collected on polytetrafluorethylene filters by using air pumps. Long-term (14 d) stationary dust samples containing airborne allergens (n = 165) were collected with electrostatic dust fall collectors (EDC). Mouse allergens were quantified by ELISA. Personnel samples were collected during bedding disposal and refilling of clean cages as well as during cage changing with and without use of cage-changing station. Animal rooms were equipped with either open cages, cages with a soft filter top, cages with a rigid filter top (static microisolation caging), or with individually ventilated cages (IVC) with either a sealed or nonsealed lid, each in positive- or negative-pressure mode. Highest personnel allergen exposure was detected during cage change and emptying of soiled cages. Allergen concentrations were lowest in rooms with sealed IVC under positive or negative pressure, with unsealed IVC under negative pressure, and with static microisolation caging. The use of cage-changing stations and a vacuum bedding-disposal system reduced median personnel exposures 14- to 25-fold, respectively. Using sealed IVC and changing stations minimized allergen exposure, indicating that state-of-the-art equipment reduces exposure to mouse allergens and decreases health risks among animal facility personnel.