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In-line digital holography is a powerful tool widely used for microscopic object imaging. Usually, in-line and out-line configurations are used to implement holographic systems, but in-line-based set-ups are preferable as they are less sensitive to mechanical vibrations and refraction index variations. However, non-desired blurred conjugate images are superposed to the reconstructed object image by using in-line systems. One strategy to remove the conjugate image contribution is to include a double-sideband filter at the Fourier plane of the system. After using the filter, data obtained at the CCD are processed to retrieve the magnitude and phase (hologram) of the diffracted wavefront while removing the conjugated image. Afterwards, a diffraction integral equation is used to digitally propagate the hologram. Despite the above-mentioned factors, there is not a thorough analysis in the literature of magnification parameters associated with the final reconstructed image, this aspect being crucial for the experimental application of the above-stated approach. Under this scenario, a theoretical analysis of the longitudinal and transverse magnifications of the reconstructed images is provided in this work. The method is validated through the simulation and experimental results of different microscopic objects: glass microspheres, a micrometric reticle, and a resolution test chart USAF 1951. The obtained results provide that the combination of magnification relations with methods for hologram propagation and optimal focused image identification is effective for object position determination. This approach could be useful for 3D microparticle localization and monitoring with optimized magnification within real-time applications. [ABSTRACT FROM AUTHOR]

In-line digital holography is a powerful tool widely used for microscopic object imaging. Usually, in-line and out-line configurations are used to implement holographic systems, but in-line-based set-ups are preferable as they are less sensitive to mechanical vibrations and refraction index variations. However, non-desired blurred conjugate images are superposed to the reconstructed object image by using in-line systems. One strategy to remove the conjugate image contribution is to include a double-sideband filter at the Fourier plane of the system. After using the filter, data obtained at the CCD are processed to retrieve the magnitude and phase (hologram) of the diffracted wavefront while removing the conjugated image. Afterwards, a diffraction integral equation is used to digitally propagate the hologram. Despite the above-mentioned factors, there is not a thorough analysis in the literature of magnification parameters associated with the final reconstructed image, this aspect being crucial for the experimental application of the above-stated approach. Under this scenario, a theoretical analysis of the longitudinal and transverse magnifications of the reconstructed images is provided in this work. The method is validated through the simulation and experimental results of different microscopic objects: glass microspheres, a micrometric reticle, and a resolution test chart USAF 1951. The obtained results provide that the combination of magnification relations with methods for hologram propagation and optimal focused image identification is effective for object position determination. This approach could be useful for 3D microparticle localization and monitoring with optimized magnification within real-time applications.