A hybrid system of holographic tomography, which utilizes rotation of a sample and two-directional, off-axis illumination is proposed. The applied type of illumination brings two major benefits. First, it offers theoretical potential for the resolution improvement with respect to conventional tomography. Second, it enables effective, numerical compensation of the defocus error, which is achieved with an accurate, noise-immune autofocusing. Hence, the main practical obstacle of hybrid tomography is removed and its high-resolution potential is put into practice. The utility of the proposed concept is experimentally demonstrated with the tomographic measurement of a photonic crystal fiber.
Exposure to laser light alters cell culture examination via optical microscopic imaging techniques based on label-free coherent digital holography. To mitigate this detrimental feature, researchers tend to use a broader spectrum and lower intensity of illumination, which can decrease the quality of holographic imaging due to lower resolution and higher noise. We study the lensless digital holographic microscopy (LDHM) ability to operate in the low photon budget (LPB) regime to enable imaging of unimpaired live cells with minimized sample interaction. Low-cost off-the-shelf components are used, promoting the usability of such a straightforward approach. We show that recording data in the LPB regime (down to 7 µW of illumination power) does not limit the contrast or resolution of the hologram phase and amplitude reconstruction compared to regular illumination. The LPB generates hardware camera shot noise, however, to be effectively minimized via numerical denoising. The ability to obtain high-quality, high-resolution optical complex field reconstruction was confirmed using the USAF 1951 amplitude sample, phase resolution test target, and finally, live glial restricted progenitor cells (as a challenging strongly absorbing and scattering biomedical sample). The proposed approach based on severely limiting the photon budget in lensless holographic microscopy method can open new avenues in high-throughout (optimal resolution, large field-of-view, and high signal-to-noise-ratio single-hologram reconstruction) cell culture imaging with minimized sample interaction.
In this Letter, we introduce an algorithm that overcomes limitations in shape measurement by holographic microscopic methods in cases of micro-optical elements with high NA, such as microlenses. The presented algorithm provides a simple method for shape reconstruction from interferometrically measured phase. The algorithm is based on the analysis of local ray transition through the measured object. We develop algorithms for holographic configurations working in transmission and reflection. The accuracy of the developed algorithms is proved by experiments and extensive simulations. We present an experiment in a holographic Mach–Zehnder configuration where we have measured and successfully reconstructed the height distribution of spherical and cylindrical microlenses with NA up to 0.3.
This paper proposes a measurement method of focusing objects with a high gradient shape of a small and large radius of curvature. The measurements are carried out on a Fourier digital holographic microscope with optimized illumination conditions maximizing the usage of the system’s numerical aperture. The obtained fringe patterns are the result of interference of deformed spherical object and spherical reference waves. The key elements of the method are the aberration compensation and calibration procedures. They provide accurate reconstruction of the object wave and determination of the focus position of the sample. The shape is calculated in two steps. First, the object wave is reconstructed at the plane of the object focus using single or multiframe phase extraction algorithm and the specialized propagation method. The step includes compensation for spherical aberration. In the second step, the sample shape is computed with the local ray approximation approach. The proposed method is experimentally validated with measurements of challenging, high gradient shapes (convex, concave) of different radiuses of curvature.