The heat distribution information of human lesions is of great value for disease analysis, diagnosis, and treatment. It is a typical inverse problem of heat conduction that deriving the distribution of internal heat sources from the temperature distribution on the body surface. This paper transforms such an inverse problem of bio-heat transfer into a direct one, thereby avoiding complex boundary conditions and regularization processes. To noninvasively reconstruct the internal heat source and its corresponding 3D temperature field in biological tissue, the adaptive simulated annealing (ASA) algorithm is used in the simulation module, where the position P(x, y, z) of point heat source in biological tissue and its corresponding temperature T are set as the optimization variables. Under a certain optimized sample, one can obtain the simulated temperature distributing on the surface of the module, then subtract the simulated temperature from the measured temperature of the same surface which was measured using a thermal infrared imager. If the sum of absolute values of the difference is smaller, it indicates that the current sample is closer to the true location and temperature of the heat source. When the values of optimization variables are determined, the corresponding 3D temperature field is also confirmed. The simulation results show the simulated position and temperature of the heat source are very approximate with those of the real experimental module. The method presented in this paper has enormous potential and promising prospects in clinical research and application, such as tumor hyperthermia, disease thermal diagnosis technology, etc.