A new thermal perfusion probe operates by imposing a thermal event on the tissue surface and directly measuring the temperature and heat flux response of the tissue with a small sensor. The thermal event is created by convectively cooling the surface with a small group of impinging jets using room temperature air. The hypothesis of this research is that this sensor can be used to provide practical burn characterization of depth and severity by determining the thickness of nonperfused tissue. To demonstrate this capability the measurement system was tested with a phantom tissue that simulates the blood perfusion of tissue. Different thicknesses of plastic were used at the surface to mimic layers of dead tissue. A mathematical model developed by Alkhwaji et al. (2012, “New Mathematical Model to Estimate Tissue Blood Perfusion, Thermal Contact Resistance and Core Temperature,” ASME J. Biomech. Eng., 134, p. 081004) is used to determine the effective values of blood perfusion, core temperature, and thermal resistance from the thermal measurements. The analytical solutions of the Pennes bioheat equation using the Green's function method is coupled with an efficient parameter estimation procedure to minimize the error between measured and analytical heat flux. Seven different thicknesses of plastic were used along with three different flow rates of perfusate to simulate burned skin of the phantom perfusion system. The resulting values of thermal resistance are a combination of the plastic resistance and thermal contact resistance between the sensor and plastic surface. Even with the uncertainty of sensor placement on the surface, the complete set of thermal resistance measurements correlate well with the layer thickness. The values are also nearly independent of the flow rate of the perfusate, which shows that the parameter estimation can successfully separate these two parameters. These results with simulated burns show the value of this minimally invasive technique to measure the thickness of nonperfused layers. This will encourage further work with this method on actual tissue burns.