3D reconstruction of unstained weakly scattering cells from a single defocused hologram

We investigate the problem of 3D complex field reconstruction corresponding to unstained red blood cells (RBCs) with a single defocused off-axis digital hologram. The main challenge in this problem is the localization of cells to the correct axial range. While investigating the volume recovery problem for a continuous phase object like the RBC, we observe an interesting feature of the backpropagated field that it does not show a clear focusing effect. Therefore, sparsity enforcement within the iterative optimization framework using a single hologram data frame cannot effectively restrict the reconstruction to the true object volume. For phase objects, it is known that the amplitude contrast of the backpropagated object field at the focus plane is minimum. We use this information available in the recovered object field in the hologram plane to device depth-dependent weights that are proportional to the inverse of amplitude contrast. This weight function is employed in the iterative steps of the optimization algorithm to assist the object volume localization. The overall reconstruction process is performed using the mean gradient descent (MGD) framework. Experimental illustrations of 3D volume reconstruction of the healthy as well as malaria-infected RBCs are presented. A test sample of polystyrene microsphere bead is also used to validate the axial localization capability of the proposed iterative technique. The proposed methodology is simple to implement experimentally and provides an approximate tomographic solution, which is axially restricted and consistent with the object field data.