To solve the inverse problem involved in fluorescence mediated tomography a regularized maximum likelihood expectation maximization (MLEM) reconstruction strategy is proposed. This technique has recently been applied to reconstruct galaxy clusters in astronomy and is adopted here. The MLEM algorithm is implemented as Richardson-Lucy (RL) scheme and includes entropic regularization and a floating default prior. Hence, the strategy is very robust against measurement noise and also avoids converging into noise patterns. Normalized Gaussian filtering with fixed standard deviation is applied for the floating default kernel. The reconstruction strategy is investigated using the XFM-2 homogeneous mouse phantom (Caliper LifeSciences Inc., Hopkinton, MA) with known optical properties. Prior to optical imaging, X-ray CT tomographic data of the phantom were acquire to provide structural context. Phantom inclusions were fit with various fluorochrome inclusions (Cy5.5) for which optical data at 60 projections over 360 degree have been acquired, respectively. Fluorochrome excitation has been accomplished by scanning laser point illumination in transmission mode (laser opposite to camera). Following data acquisition, a 3D triangulated mesh is derived from the reconstructed CT data which is then matched with the various optical projection images through 2D linear interpolation, correlation and Fourier transformation in order to assess translational and rotational deviations between the optical and CT imaging systems. Preliminary results indicate that the proposed regularized MLEM algorithm, when driven with a constant initial condition, yields reconstructed images that tend to be smoother in comparison to classical MLEM without regularization. Once the floating default prior is included this bias was significantly reduced.