An automatic method that fully compensates the quantitative phase measurements in off-axis digital holographic microscopy (DHM) is presented. The two main perturbations of the quantitative phase measurements in off-axis DHM are automatically removed. While the curvature phase flaw introduced by the microscope objective is avoided by the use of an optimized telecentric imaging system for the recording of the holograms, the remaining phase perturbation due to the tilt of the reference wave is removed by the automatic computation of a digital compensating reference wave. The method has been tested on both nonbiological and biological samples with and improving on the quality of the recovered phase maps.
The description and validation of an ImageJ open-source plugin to numerically simulate and reconstruct digital lensless holographic microscopy (DLHM) holograms are presented. Two modules compose the presented plugin: the simulation module implements a discrete version of the Rayleigh–Somerfield diffraction formula, which allows the user to directly build a simulated hologram from a known phase and/or amplitude object by just introducing the geometry parameters of the simulated setup; the plugin’s reconstruction module implements a discrete version of the Kirchhoff–Helmholtz diffraction integral, thus allowing the user to reconstruct DLHM holograms by introducing the parameters of the acquisition setup and the desired reconstruction distance. The plugin offers the two said modules within the robust environment provided by a complete set of built-in tools for image processing available in ImageJ. While the simulation module has been validated through the evaluation of the forecasted lateral resolution of a DLHM setup in terms of the numerical aperture, the reconstruction module is tested by means of reconstructing experimental DLHM holograms of biological samples.
An ImageJ plugin for numerical wave propagation is presented. The plugin provides ImageJ, the well-known software for image processing, with the capability of computing numerical wave propagation by the use of angular spectrum, Fresnel, and Fresnel-Bluestein algorithms. The plugin enables numerical wave propagation within the robust environment provided by the complete set of built-in tools for image processing available in ImageJ. The plugin can be used for teaching and research purposes. We illustrate its use to numerically recreate Poisson's spot and Babinet's principle, and in the numerical reconstruction of digitally recorded holograms from millimeter-sized and pure phase microscopic objects.
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