In this chapter we show how digital holography (DH) can be efficiently used as a free and noninvasive investigation tool capable of performing quantitative and qualitative mapping of biological samples. A detailed description of the recent methods based on the possibility offered by DH to numerically manage the reconstructed wavefronts is reported. Depending on the sample and on the investigation to be performed it is possible, by a single acquired image, to recover information on the optical path length changes and, choosing the more suitable numerical method, to obtain quantitative phase map distributions. The progress achieved in the reconstruction methods will certainly find useful applications in the field of the biological analysis.