Based on the finite element analysis, this study applies three iterative regularization algorithms to the 3D reconstruction of fluorescent yield, including conjugate gradient least square (CGLS), least square QR decomposition (LSQR), and 2-order pre-iteration method. By using a non-contact, multi-angle transmission imaging system, the experiments of single fluorescent target and double targets with the common-used cylinder phantom are conducted. Experimental results show that the above three methods could estimate the position of fluorescence targets accurately, while the time cost of each method is only about 2% of that of algebraic reconstruction technology (ART).