Weighted filtered backprojection for quantitative fluorescence optical projection tomography

Abstract

Reconstructing images from a set of fluorescence optical projection tomography (OPT) projections is a relatively new problem. Several physical aspects of fluorescence OPT necessitate a different treatment of the inverse problem to that required for non-fluorescence tomography. Given a fluorophore within the depth of field of the imaging system, the power received by the optical system, and therefore the CCD detector, is related to the distance of the fluorophore from the objective entrance pupil. Additionally, due to the slight blurring of images of sources positioned off the focal plane, the CCD image of a fluorophore off the focal plane is lower in intensity than the CCD image of an identical fluorophore positioned on the focal plane. The filtered backprojection (FBP) algorithm does not take these effects into account and so cannot be expected to yield truly quantitative results. A full model of image formation is introduced which takes into account the effects of isotropic emission and defocus. The model is used to obtain a weighting function which is used in a variation of the FBP algorithm called weighted filtered backprojection (WFBP). This new algorithm is tested with simulated data and with experimental data from a phantom consisting of fluorescent microspheres embedded in an agarose gel.