Between welcome culture and border fence: A dataset on the European refugee crisis in German newspaper reports.

Newspaper reports provide a rich source of information on the unfolding of public debates, which can serve as basis for inquiry in political science. Such debates are often triggered by critical events, which attract public attention and incite the reactions of political actors: crisis sparks the debate. However, due to the challenges of reliable annotation and modeling, few large-scale datasets with high-quality annotation are available. This paper introduces DebateNet2.0, which traces the political discourse on the 2015 European refugee crisis in the German quality newspaper taz. The core units of our annotation are political claims (requests for specific actions to be taken) and the actors who advance them (politicians, parties, etc.). Our contribution is twofold. First, we document and release DebateNet2.0 along with its companion R package, mardyR. Second, we outline and apply a Discourse Network Analysis (DNA) to DebateNet2.0, comparing two crucial moments of the policy debate on the "refugee crisis": the migration flux through the Mediterranean in April/May and the one along the Balkan route in September/October. We guide the reader through the methods involved in constructing a discourse network from a newspaper, demonstrating that there is not one single discourse network for the German migration debate, but multiple ones, depending on the research question through the associated choices regarding political actors, policy fields and time spans.

Directional Aspects of Vegetation Linear and Circular Polarization Biosignatures.

Homochirality is a generic and unique property of all biochemical life and it is considered a universal and agnostic biosignature. Upon interaction with unpolarized light, homochirality induces fractional circular polarization in the light that is scattered from it, which can be sensed remotely. As such, it can be a prime candidate biosignature in the context of future life detection missions and observatories. The linear polarizance of vegetation is also sometimes envisaged as a biosignature, although it does not share the same molecular origin as circular polarization. It is known that linear polarization of surfaces is strongly dependent on the phase angle. The relationship between the phase angle and circular polarization stemming from macromolecular assemblies such as in vegetation, however, remained unclear. In this study, using the average of 27 different species, we demonstrate that the circular polarization-phase angle dependency of vegetation induces relatively small changes in spectral shape and mostly affects the signal magnitude. With these results, we underline the use of circular spectropolarimetry as a promising agnostic biosignature complementary to the use of linear spectropolarimetry and scalar reflectance.

Active focal-plane coronagraphy with liquid-crystal spatial-light modulators: broadband contrast performance in the visible.

The technological progress in spatial-light modulator (SLM) technology has made it possible to use those devices as programmable active focal-plane phase coronagraphic masks, opening the door to novel versatile and adaptive high-contrast imaging observation strategies. However, the scalar nature of the SLM-induced phase response is a potential hurdle when applying the approach to wideband light, as is typical in astronomical imaging. For the first time, to our knowledge, we present laboratory results with broadband light (up to ∼12% bandwidth) for two commercially available SLM devices used as active focal-plane phase masks in the visible regime (640 nm). It is shown that under ideal or realistic telescope aperture conditions, the contrast performance is negligibly affected in this bandwidth regime, reaching a sufficient level for ground-based high-contrast imaging, which is typically dominated by atmospheric residuals.

Implementing focal-plane phase masks optimized for real telescope apertures with SLM-based digital adaptive coronagraphy.

Direct imaging of exoplanets or circumstellar disk material requires extreme contrast at the 10-6 to 10-12 levels at < 100 mas angular separation from the star. Focal-plane mask (FPM) coronagraphic imaging has played a key role in this field, taking advantage of progress in Adaptive Optics on ground-based 8 + m class telescopes. However, large telescope entrance pupils usually consist of complex, sometimes segmented, non-ideal apertures, which include a central obstruction for the secondary mirror and its support structure. In practice, this negatively impacts wavefront quality and coronagraphic performance, in terms of achievable contrast and inner working angle. Recent theoretical works on structured darkness have shown that solutions for FPM phase profiles, optimized for non-ideal apertures, can be numerically derived. Here we present and discuss a first experimental validation of this concept, using reflective liquid crystal spatial light modulators as adaptive FPM coronagraphs.

Improved Tracking and Resolution of Bacteria in Holographic Microscopy Using Dye and Fluorescent Protein Labeling.

Digital holographic microscopy (DHM) is an emerging imaging technique that permits instantaneous capture of a relatively large sample volume. However, large volumes usually come at the expense of lower spatial resolution, and the technique has rarely been used with prokaryotic cells due to their small size and low contrast. In this paper we demonstrate the use of a Mach-Zehnder dual-beam instrument for imaging of labeled and unlabeled bacteria and microalgae. Spatial resolution of 0.3 µm is achieved, providing a sampling of several pixels across a typical prokaryotic cell. Both cellular motility and morphology are readily recorded. The use of dyes provides both amplitude and phase contrast improvement and is of use to identify cells in dense samples.

Robust, compact implementation of an off-axis digital holographic microscope.

Recent advances in digital technologies, such as high-speed computers and large-format digital imagers, have led to a burgeoning interest in the science and engineering of digital holographic microscopy (DHM). Here we report on a novel off-axis DHM, based on a twin-beam optical design, which avoids the limitations of prior systems, and provides many advantages, including compactness, intrinsic stability, robustness against misalignment, ease of use, and cost. These advantages are traded for a physically constrained sample volume, as well as a fixed fringe spacing. The first trade is not overly restrictive for most applications, and the latter provides for a pre-set assembly alignment that optimizes the spatial frequency sampling. Moreover, our new design supports use in both routine laboratory settings as well as extreme environments without any sacrifice in performance, enabling ready observation of microbial species in the field. The instrument design is presented in detail here, along with a demonstration of bacterial video imaging at sub-micrometer resolution at temperatures down to -15 °C.

A Mach-Zender digital holographic microscope with sub-micrometer resolution for imaging and tracking of marine micro-organisms.

Digital holographic microscopy is an ideal tool for investigation of microbial motility. However, most designs do not exhibit sufficient spatial resolution for imaging bacteria. In this study we present an off-axis Mach-Zehnder design of a holographic microscope with spatial resolution of better than 800 nm and the ability to resolve bacterial samples at varying densities over a 380 µm × 380 µm × 600 µm three-dimensional field of view. Larger organisms, such as protozoa, can be resolved in detail, including cilia and flagella. The instrument design and performance are presented, including images and tracks of bacterial and protozoal mixed samples and pure cultures of six selected species. Organisms as small as 1 µm (bacterial spores) and as large as 60 µm (Paramecium bursaria) may be resolved and tracked without changes in the instrument configuration. Finally, we present a dilution series investigating the maximum cell density that can be imaged, a type of analysis that has not been presented in previous holographic microscopy studies.

Measurement of absolute cell volume, osmotic membrane water permeability, and refractive index of transmembrane water and solute flux by digital holographic microscopy.

A dual-wavelength digital holographic microscope to measure absolute volume of living cells is proposed. The optical setup allows us to reconstruct two quantitative phase contrast images at two different wavelengths from a single hologram acquisition. When adding the absorbing dye fast green FCF as a dispersive agent to the extracellular medium, cellular thickness can be univocally determined in the full field of view. In addition to the absolute cell volume, the method can be applied to derive important biophysical parameters of living cells including osmotic membrane water permeability coefficient and the integral intracellular refractive index (RI). Further, the RI of transmembrane flux can be determined giving an indication about the nature of transported solutes. The proposed method is applied to cultured human embryonic kidney cells, Chinese hamster ovary cells, human red blood cells, mouse cortical astrocytes, and neurons.

Label-free cytotoxicity screening assay by digital holographic microscopy.

We introduce a label-free technology based on digital holographic microscopy (DHM) with applicability for screening by imaging, and we demonstrate its capability for cytotoxicity assessment using mammalian living cells. For this first high content screening compatible application, we automatized a digital holographic microscope for image acquisition of cells using commercially available 96-well plates. Data generated through both label-free DHM imaging and fluorescence-based methods were in good agreement for cell viability identification and a Z'-factor close to 0.9 was determined, validating the robustness of DHM assay for phenotypic screening. Further, an excellent correlation was obtained between experimental cytotoxicity dose-response curves and known IC50 values for different toxic compounds. For comparable results, DHM has the major advantages of being label free and close to an order of magnitude faster than automated standard fluorescence microscopy.

Accelerated autofocusing of off-axis holograms using critical sampling.

In this Letter we propose a fast off-axis hologram autofocusing (AF) approach that is based on the redundant data elimination by the critical resampling of the contained complex field. Implementation of the proposed methodology enables the real-time AF with up to 12× speed-up factors in comparison to the classical approach. The method is further extended for single-shot physical autofocus of the fluorescence imaging channel of multimodal imaging instruments capable of off-axis hologram acquisition.

Enhanced robustness digital holographic microscopy for demanding environment of space biology.

We describe an optimized digital holographic microscopy system (DHM) suitable for high-resolution visualization of living cells under conditions of altered macroscopic mechanical forces such as those that arise from changes in gravitational force. Experiments were performed on both a ground-based microgravity simulation platform known as the random positioning machine (RPM) as well as during a parabolic flight campaign (PFC). Under these conditions the DHM system proved to be robust and reliable. In addition, the stability of the system during disturbances in gravitational force was further enhanced by implementing post-processing algorithms that best exploit the intrinsic advantages of DHM for hologram autofocusing and subsequent image registration. Preliminary results obtained in the form of series of phase images point towards sensible changes of cytoarchitecture under states of altered gravity.

Early cell death detection with digital holographic microscopy.

BACKGROUND: Digital holography provides a non-invasive measurement of the quantitative phase shifts induced by cells in culture, which can be related to cell volume changes. It has been shown previously that regulation of cell volume, in particular as it relates to ionic homeostasis, is crucially involved in the activation/inactivation of the cell death processes. We thus present here an application of digital holographic microscopy (DHM) dedicated to early and label-free detection of cell death. METHODS AND FINDINGS: We provide quantitative measurements of phase signal obtained on mouse cortical neurons, and caused by early neuronal cell volume regulation triggered by excitotoxic concentrations of L-glutamate. We show that the efficiency of this early regulation of cell volume detected by DHM, is correlated with the occurrence of subsequent neuronal death assessed with the widely accepted trypan blue method for detection of cell viability. CONCLUSIONS: The determination of the phase signal by DHM provides a simple and rapid optical method for the early detection of cell death.

Extended depth-of-focus by digital holographic microscopy.

A recurrent problem in microscopy is the finite depth-of-focus linked to the NA of microscope objectives. Digital holographic microscopy (DHM) has the unique feature of being able to numerically change the focus from a single hologram without the need of moving the sample. Extended depth of focus of amplitude images has been demonstrated, but it has marginal interest for the metrological application of DHM that needs the topography. In this Letter, we demonstrate that DHM is able to provide not only extended depth-of-focus amplitude images but extended focused complex data from which the topography is computed. For this purpose, reflection and transmission measurements on micro-optics (microlens and retroreflector) performed by using standard reconstruction or the extended focused complex data are compared. These experiments demonstrate that DHM measures, from a single hologram acquisition, the accurate sample topography on a numerically increased depth-of-focus.

Digital holographic microscopy real-time monitoring of cytoarchitectural alterations during simulated microgravity.

Previous investigations on mammalian cells have shown that microgravity, either that experienced in space, or simulated on earth, causes severe cellular modifications that compromise tissue determination and function. The aim of this study is to investigate, in real time, the morphological changes undergone by cells experiencing simulated microgravity by using digital holographic microscopy (DHM). DHM analysis of living mouse myoblasts (C2C12) is undertaken under simulated microgravity with a random positioning machine. The DHM analysis reveals cytoskeletal alterations similar to those previously reported with conventional methods, and in agreement with conventional brightfield fluorescence microscopy a posteriori investigation. Indeed, DHM is shown to be able to noninvasively and quantitatively detect changes in actin reticular formation, as well as actin distribution, in living unstained samples. Such results were previously only obtainable with the use of labeled probes in conjunction with conventional fluorescence microscopy, with all the classically described limitations in terms of bias, bleaching, and temporal resolution.

Digital holographic reflectometry.

Digital holographic microscopy (DHM) is an interferometric technique that allows real-time imaging of the entire complex optical wavefront (amplitude and phase) reflected by or transmitted through a sample. To our knowledge, only the quantitative phase is exploited to measure topography, assuming homogeneous material sample and a single reflection on the surface of the sample. In this paper, dual-wavelength DHM measurements are interpreted using a model of reflected wave propagation through a three-interfaces specimen (2 layers deposited on a semi-infinite layer), to measure simultaneously topography, layer thicknesses and refractive indices of micro-structures. We demonstrate this DHM reflectometry technique by comparing DHM and profilometer measurement of home-made SiO(2)/Si targets and Secondary Ion Mass Spectrometry (SIMS) sputter craters on specimen including different multiple layers.

Cell morphology and intracellular ionic homeostasis explored with a multimodal approach combining epifluorescence and digital holographic microscopy.

The authors have developed a live-cell multimodality microscope combining epifluorescence with digital holographic microscopy; it has been implemented with a decoupling procedure allowing to separately measure from the quantitative phase important cell parameters including absolute volume, shape and integral intracellular refractive index. In combination with the numerous different specific fluorescent cellular probes, this multimodality microscopy can address important issues in cell biology. This is demonstrated by the study of intracellular calcium homeostasis associated with the change in cell volume, which play a critical role in the excitotoxicity-induced neuronal death.

Endoscopic low-coherence topography measurement for upper airways and hollow samples.

To evaluate the severity of airway pathologies, quantitative dimensioning of airways is of utmost importance. Endoscopic vision gives a projective image and thus no true scaling information can be directly deduced from it. In this article, an approach based on an interferometric setup, a low-coherence laser source and a standard rigid endoscope is presented, and applied to hollow samples measurements. More generally, the use of the low-coherence interferometric setup detailed here could be extended to any other endoscopy-related field of interest, e.g., gastroscopy, arthroscopy and other medical or industrial applications where tri-dimensional topology is required. The setup design with a multiple fibers illumination system is presented. Demonstration of the method ability to operate on biological samples is assessed through measurements on ex vivo pig bronchi.

Suppression of the zero-order term in off-axis digital holography through nonlinear filtering.

We present experimental validation of a new reconstruction method for off-axis digital holographic microscopy (DHM). This method effectively suppresses the object autocorrelation, namely, the zero-order term, from holographic data, thereby improving the reconstruction bandwidth of complex wavefronts. The algorithm is based on nonlinear filtering and can be applied to standard DHM setups with realistic recording conditions. We study the robustness of the technique under different experimental configurations, and quantitatively demonstrate its enhancement capabilities on phase signals.

Digital holographic microscopy investigation of second harmonic generated at a glass/air interface.

Optical second-harmonic generation, thanks to its coherent nature, is a suitable signal for interferometric measurements such as digital holography, a well-established imaging technique that allows recovery of complex diffraction wave fields from which it is possible to extract both amplitude-contrast and quantitative phase images. Here, we report on a multifunctional form of microscopy, namely, second-harmonic generation digital holographic microscopy. As a proof of concept, we have investigated the second-harmonic signal generated at the glass/air interface of a microscope slide under focused femtosecond laser illumination, and we propose, for the first time to our knowledge, a representation and interpretation of the recovered phase. In this simple yet educative case study, we observe that the second harmonic is generated by the axial component of the incident field polarization.

Noninvasive characterization of the fission yeast cell cycle by monitoring dry mass with digital holographic microscopy.

Digital holography microscopy (DHM) is an optical technique which provides phase images yielding quantitative information about cell structure and cellular dynamics. Furthermore, the quantitative phase images allow the derivation of other parameters, including dry mass production, density, and spatial distribution. We have applied DHM to study the dry mass production rate and the dry mass surface density in wild-type and mutant fission yeast cells. Our study demonstrates the applicability of DHM as a tool for label-free quantitative analysis of the cell cycle and opens the possibility for its use in high-throughput screening.