In-line Digital holographic microscopy is easy to implement because it only requires a coherent illumination source. An out-of-focus image (hologram) of the sample is recorded and numerically reconstructed to retrieve phase shift information due to the sample. Effects of the optical setup, such as aberrations, can distort the hologram signal, and ignoring these effects results in biased reconstructions. In this context, we study the effects of misalignment and wrong cover-slip thickness by statistically analysing the reconstructions of beads distributed in the whole field of view. Our aberration-wise reconstruction approach produces debiased estimations and allows precise and realistic estimation of optical aberrations from a single hologram.
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