An optical sensor and a method of operating the optical sensor are provided. The optical sensor includes a light source configured to emit a light, and a path adjuster configured to adjust a traveling path of the light to reflect the light at a first time, and allow the light to pass through the path adjuster at a second time. The optical sensor further includes a light receiver configured to receive a reference light among the reflected light, and receive, among the light passing through the path adjuster, a measurement light related to a target material.