Abstract. In the last few years, classification of cells by machine learning has become frequently used in biology. However, most of the approaches are based on morphometric (MO) features, which are not quantitative in terms of cell mass. This may result in poor classification accuracy. Here, we study the potential contribution of coherence-controlled holographic microscopy enabling quantitative phase imaging for the classification of cell morphologies. We compare our approach with the commonly used method based on MO features. We tested both classification approaches in an experiment with nutritionally deprived cancer tissue cells, while employing several supervised machine learning algorithms. Most of the classifiers provided higher performance when quantitative phase features were employed. Based on the results, it can be concluded that the quantitative phase features played an important role in improving the performance of the classification. The methodology could be valuable help in refining the monitoring of live cells in an automated fashion. We believe that coherencecontrolled holographic microscopy, as a tool for quantitative phase imaging, offers all preconditions for the accurate automated analysis of live cell behavior while enabling noninvasive label-free imaging with sufficient contrast and high-spatiotemporal phase sensitivity.
Coherence-controlled holographic microscopy (CCHM) is a realtime, wide-field, and quantitative light-microscopy technique enabling 3D imaging of electromagnetic fields, providing complete information about both their intensity and phase. These attributes make CCHM a promising candidate for performance assessment of phase-altering metasurfaces, a new class of artificial materials that allow to manipulate the wavefront of passing light and thus provide unprecedented functionalities in optics and nanophotonics. In this paper, we report on our investigation of phase imaging of plasmonic metasurfaces using holographic microscopy. We demonstrate its ability to obtain phase information from the whole field of view in a single measurement on a prototypical sample consisting of silver nanodisc arrays. The experimental data were validated using FDTD simulations and a theoretical model that relates the obtained phase image to the optical response of metasurface building blocks. Finally, in order to reveal the full potential of CCHM, we employed it in the analysis of a simple metasurface represented by a plasmonic zone plate. By scanning the sample along the optical axis we were able to create a quantitative 3D phase map of fields transmitted through the zone plate. The presented results prove that CCHM is inherently suited to the task of metasurface characterization. Moreover, as the temporal resolution is limited only by the camera framerate, it can be even applied in analysis of actively tunable metasurfaces.
Light vortices carry orbital angular momentum and have a variety of applications in optical manipulation, high-capacity communications or microscopy. Here we propose a new concept of full-field vortex topographic microscopy enabling a reference-free displacement and shape measurement of reflective samples. The sample surface is mapped by an array of light spots enabling quantitative reconstruction of the local depths from defocused wavefronts. Light from the spots is converted to a lattice of mutually uncorrelated double-helix point spread functions (PSFs) whose angular rotation enables depth estimation. The PSFs are created by self-interference of optical vortices that originate from the same wavefront and are shaped by a spiral phase mask (SPM). The method benefits from the isoplanatic PSFs whose shape and size remain unchanged under defocusing, ensuring high precision in a wide range of measured depths. The technique was tested using a microscope Nikon Eclipse E600 working with a micro-hole plate providing structured illumination and the SPM placed in the imaging path. The depth measurement was demonstrated in the range of 11 µm exceeding the depth of field of the microscope objective up to 19 times. Throughout this range, the surface depth was mapped with the precision better than 30 nm at the lateral positions given with the precision better than 10 nm. Application potential of the method was demonstrated by profiling the top surface of a bearing ball and reconstructing the three-dimensional relief of a reflection phase grating.
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