Lensless holographic microscope (LHM) is an emerging very promising technology that provides high-quality imaging and analysis of biological samples without utilizing any lens for imaging. Due to its small size and reduced price, LHM can be a very useful tool for the point-of-care diagnosis of diseases, sperm assessment, or microfluidics, among others, not only employed in advanced laboratories but also in poor and/or remote areas. Recently, several LHMs have been reported in the literature. However, complete characterization of their optical parameters remains not much presented yet. Hence, we present a complete analysis of the performance of a compact, reduced cost, and high-resolution LHM. In particular, optical parameters such as lateral and axial resolutions, lateral magnification, and field of view are discussed into detail, comparing the experimental results with the expected theoretical values for different layout configurations. We use high-resolution amplitude and phase test targets and several microbeads to characterize the proposed microscope. This characterization is used to define a balanced and matched setup showing a good compromise between the involved parameters. Finally, such a microscope is utilized for visualization of static, as well as dynamic biosamples.

Quantitative phase imaging (QPI) is nowadays a powerful tool for visualization and analysis of biological processes. QPI is usually attained from specifically designed optical microscopes retrieving phase information in a quantitative way. In this paper we report on an extremely simple, low cost and compact way to update a standard bright-field microscope with coherent sensing capabilities. It is based on the in-line Gabor holography concept and only needs to replace the illumination broadband source of the regular microscope with a coherent one. The proposed methodology is completed by the recording of a digital in-line Gabor hologram instead of regular imaging conditions and by the numerical processing of the recorded hologram to finally achieve QPI. The selection of the defocus distance is critical to finally achieve high quality phase imaging, so calibration considering phase and amplitude test targets is presented for the proper definition of such defocus distance. In addition, the selected configuration is experimentally validated using different samples (microbeads, cheek cells and alive spermatozoa). All the experiments are implemented in a regular Olympus BX-60 upright microscope using a 20X 0.46NA objective lens. The proposed method shows how to convert a regular microscope into a holographic one with probably the minimum needed modifications and with the main limitation coming from the Gabor imaging conditions (weak diffractive samples and twin image presence).

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