Novel biophotonic phantoms are provided herein that can accurately mimic the optical properties of living tissue. The disclosed biophotonic phantoms comprise hemoglobin (Hb) in a native conformation that is distributed in a solid polymer matrix. Methods of producing the disclosed biophotonic phantoms are also provided. The biophotonic phantoms can be used, for example, to calibrate or test an optical imaging system, such as a near infrared spectroscopy imaging system.