A new optical tissue phantom is presented, which consists of a polymer film containing scatterers and absorbers. With this tissue phantom, tissue models can be constructed having the optical properties and layered structure similar to those in living tissue. Optical properties can be specified for each layer, with a resolution of 20 micrometers . With this tissue phantom, a new laser Doppler testing principle is developed using a repetitive construction of static and moving layers. Monte Carlo simulations show that the requirements to the optical matching quality provided by the medium between the layers are not very restrictive. Two applications of the new tester are shown. For a single laser Doppler blood flow monitor, the depth sensitivity characteristics of two fiber optic probes are measured. Furthermore, an attempt is made to compare the outputs of two different laser Doppler instruments measuring on the same tissue phantom, with the same flow situation. Such a comparison may be important when new instruments are used for a specific medical application.© (1998) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.