The optical properties of human tissues are an important parameter in medical diagnostics and therapy. The knowledge of these parameters can encourage the development of automated, computer-driven optical tissue analysis methods. We determine the absorption coefficient μ_a and scattering coefficient μs′ of different tissue types obtained during parotidectomy in the wavelength range of 250 to 800 nm. These values are determined by high precision integrating sphere measurements in combination with an optimized inverse Monte Carlo simulation. To conserve the optical behavior of living tissues, the optical spectroscopy measurements are performed immediately after tissue removal. Our study includes fresh samples of the ear, nose, and throat (ENT) region, as muscle tissue, nervous tissue, white adipose tissue, stromal tissue, parotid gland, and tumorous tissue of five patients. The measured behavior of adipose corresponds well with the literature, which sustains the applied method. It is shown that muscle is well supplied with blood as it features the same characteristic peaks at 430 and 555 nm in the absorption curve. The parameter μs′ decreases for all tissue types above 570 nm. The accuracy is adequate for the purposes of providing μ_a and μs′ of different human tissue types as muscle, fat, nerve, or gland tissue, which are embedded in large complex structures such as in the ENT area. It becomes possible for the first time to present reasonable results for the optical behavior of human soft tissue located in the ENT area and in the near-UV, visual, and near-infrared areas.