Self-interference digital holography (SIDH) has the potential to achieve imaging of biological objects over large axial ranges from two-dimensional images. Combined with single molecule localization microscopy, SIDH can localize fluorescent objects with near nanometer precision over a wide axial range without using any mechanical focusing. Using these techniques, fluorescence microscopy has enabled imaging below the diffraction limit, opening a new path to super-resolution microscopy. However, the high sensibility to background noise degrades rapidly the localization ability. Therefore, it is necessary to enhance the signal-to-noise ratio. Striving for that, one approach consists in improving the light efficiency of SIDH. So, based on a large amount of numerical simulations, this paper presents an efficient methodology to analyze and optimize the SIDH performance for better photon emission utilization across a large axial range. Simulations are completed with the results of experiments conducted on fluorescent microspheres, which compare performances of the SIDH system using an optimized and a non-optimized point-spread hologram and fully demonstrate the validity of the methodology presented by the authors. Finally, the three-dimensional imaging capability of the optimized SIDH system is assessed with a last experimentation were small particles emitting low numbers of photons are imaged over a wide axial range.

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