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Optical tomography has emerged as a non-invasive imaging method, providing three-dimensional insights into subcellular structures and thereby enabling a deeper understanding of cellular functions, interactions, and processes. Conventional optical tomography methods are constrained by a limited illumination scanning range, leading to anisotropic resolution and incomplete imaging of cellular structures. To overcome this problem, we employ a compact multi-core fibre-optic cell rotator system that facilitates precise optical manipulation of cells within a microfluidic chip, achieving full-angle projection tomography with isotropic resolution. Moreover, we demonstrate an AI-driven tomographic reconstruction workflow, which can be a paradigm shift from conventional computational methods, often demanding manual processing, to a fully autonomous process. The performance of the proposed cell rotation tomography approach is validated through the three-dimensional reconstruction of cell phantoms and HL60 human cancer cells. The versatility of this learning-based tomographic reconstruction workflow paves the way for its broad application across diverse tomographic imaging modalities, including but not limited to flow cytometry tomography and acoustic rotation tomography. Therefore, this AI-driven approach can propel advancements in cell biology, aiding in the inception of pioneering therapeutics, and augmenting early-stage cancer diagnostics., Comment: 15 pages, 6 figures

Real-time deformability cytometry (RT-DC) is an established method that quantifies features like size, shape, and stiffness for whole cell populations on a single-cell level in real time. To extract the cell stiffness, a lookup table (LUT) disentangles the experimentally derived steady state cell deformation and the projected area, yielding the Young's modulus. So far, two lookup tables exists, but are limited to simple linear material models and cylindrical channel geometries. Here, we present two new lookup tables for RT-DC based on a neo-Hookean hyperelastic material numerically derived by simulations based on the finite element method in square and cylindrical channel geometries. At the same time, we quantify the influence of the shear-thinning behaviour of the surrounding medium on the stationary deformation of cells in RT-DC and discuss the applicability and impact of the proposed LUTs regarding past and future RT-DC data analysis. Additionally, we provide insights about the cell strain and stresses, as well as the influence resulting from the rotational symmetric assumption on the cell deformation and volume estimation. The new lookup tables as well as the numerical cell shapes are made freely available.

Quantitative phase imaging (QPI) is a label-free technique providing both morphology and quantitative biophysical information in biomedicine. However, applying such a powerful technique to in vivo pathological diagnosis remains challenging. Multi-core fiber bundles (MCFs) enable ultra-thin probes for in vivo imaging, but current MCF imaging techniques are limited to amplitude imaging modalities. We demonstrate a computational lensless microendoscope that uses an ultra-thin bare MCF to perform quantitative phase imaging of biomedical samples with up to 1 {\mu}m lateral resolution and nanoscale axial resolution. The incident complex light field at the measurement side is precisely reconstructed from a single-shot far-field speckle pattern at the detection side, enabling digital focusing and 3D volumetric reconstruction without any mechanical movement. The accuracy of the quantitative phase reconstruction is validated by imaging the phase target and hydrogel beads through the MCF. With the proposed imaging modality, 3D imaging of human cancer cells is achieved through the ultra-thin fiber endoscope, promising widespread clinical applications., Comment: 16pages, 6 figures

Despite fluorescent cell-labelling being widely employed in biomedical studies, some of its drawbacks are inevitable, with unsuitable fluorescent probes or probes inducing a functional change being the main limitations. Consequently, the demand for and development of label-free methodologies to classify cells is strong and its impact on precision medicine is relevant. Towards this end, high-throughput techniques for cell mechanical phenotyping have been proposed to get a multidimensional biophysical characterization of single cells. With this motivation, our goal here is to investigate the extent to which an unsupervised machine learning methodology, which is applied exclusively on morpho-rheological markers obtained by real-time deformability and fluorescence cytometry (RT-FDC), can address the difficult task of providing label-free discrimination of reticulocytes from mature red blood cells. We focused on this problem, since the characterization of reticulocytes (their percentage and cellular features) in the blood is vital in multiple human disease conditions, especially bone-marrow disorders such as anemia and leukemia. Our approach reports promising label-free results in the classification of reticulocytes from mature red blood cells, and it represents a step forward in the development of high-throughput morpho-rheological-based methodologies for the computational categorization of single cells. Besides, our methodology can be an alternative but also a complementary method to integrate with existing cell-labelling techniques., Comment: 13 pages, 3 figures, 4 tables

The shortest path (SP) and shortest paths tree (SPT) problems arise both as direct applications and as subroutines of overlay algorithms solving more complex problems such as the constrained shortest path (CSP) or the constrained minimum Steiner tree (CMST) problems. Often, such algorithms do not use the result of an SP subroutine if its total cost is greater than a given bound. For example, for delay-constrained problems, paths resulting from a least-delay SP run and whose delay is greater than the delay constraint of the original problem are not used by the overlay algorithm to construct its solution. As a result of the existence of these bounds, and because the Dijkstra SP algorithm discovers paths in increasing order of cost, we can terminate the SP search earlier, i.e., once it is known that paths with a greater total cost will not be considered by the overlay algorithm. This early termination allows to reduce the runtime of the SP subroutine, thereby reducing the runtime of the overlay algorithm without impacting its final result. We refer to this adaptation of Dijkstra for centralized implementations as bounded Dijkstra (BD). On the example of CSP algorithms, we confirm the usefulness of BD by showing that it can reduce the runtime of some algorithms by 75% on average., Comment: 9 pages -- accompanying web page: https://lora.lkn.ei.tum.de

State synchronisation in clustered Software Defined Networking controller deployments ensures that all instances of the controller have the same state information in order to provide redundancy. Current implementations of controllers use a strong consistency model, where configuration changes must be synchronised across a number of instances before they are applied on the network infrastructure. For large deployments, this blocking process increases the delay of state synchronisation across cluster members and consequently has a detrimental effect on network operations that require rapid response, such as fast failover and Quality of Service applications. In this paper, we introduce an adaptive consistency model for SDN Controllers that employs concepts of eventual consistency models along with a novel `cost-based' approach where strict synchronisation is employed for critical operations that affect a large portion of the network resources while less critical changes are periodically propagated across cluster nodes. We use simulation to evaluate our model and demonstrate the potential gains in performance., Comment: 7 pages

The routing algorithms used by current operators aim at coping with the demanded QoS requirements while optimizing the use of their network resources. These algorithms rely on the optimal substructure property (OSP), which states that an optimal path contains other optimal paths within it. However, we show that QoS metrics such as queuing delay and buffer consumption do not satisfy this property, which implies that the used algorithms lose their optimality and/or completeness. This negatively impacts the operator economy by causing a waste of network resources and/or violating Service Level Agreements (SLAs). In this paper, we propose a new so-called Mn taxonomy defining new metric classes. An Mn metric corresponds to a metric which requires the knowledge of the n previously traversed edges to compute its value at a given edge. Based on this taxonomy, we present three solutions for solving routing problems with the newly defined classes of metrics. We show that state-of-the-art algorithms based on the OSP indeed lose their original optimality and/or completeness properties while our proposed solutions do not, at the price of an increased computation time., Comment: 2018 International Conference on Communications (ICC 2018), Kansas City, MO (USA)

In a recent study entitled "Cell nuclei have lower refractive index and mass density than cytoplasm", we provided strong evidence indicating that the nuclear refractive index (RI) is lower than the RI of the cytoplasm for several cell lines. In a complementary study in 2017, entitled "Is the nuclear refractive index lower than cytoplasm? Validation of phase measurements and implications for light scattering technologies", Steelman et al. observed a lower nuclear RI also for other cell lines and ruled out methodological error sources such as phase wrapping and scattering effects. Recently, Yurkin composed a comment on these 2 publications, entitled "How a phase image of a cell with nucleus refractive index smaller than that of the cytoplasm should look like?", putting into question the methods used for measuring the cellular and nuclear RI in the aforementioned publications by suggesting that a lower nuclear RI would produce a characteristic dip in the measured phase profile in situ. We point out the difficulty of identifying this dip in the presence of other cell organelles, noise, or blurring due to the imaging point spread function. Furthermore, we mitigate Yurkin's concerns regarding the ability of the simple-transmission approximation to compare cellular and nuclear RI by analyzing a set of phase images with a novel, scattering-based approach. We conclude that the absence of a characteristic dip in the measured phase profiles does not contradict the usage of the simple-transmission approximation for the determination of the average cellular or nuclear RI. Our response can be regarded as an addition to the response by Steelman, Eldridge and Wax. We kindly ask the reader to attend to their thorough ascertainment prior to reading our response., Comment: 5 pages, 3 figures, response to comment

In this chapter, we introduce digital holographic microscopy (DHM) as a marker-free method to determine the refractive index of single, spherical cells in suspension. The refractive index is a conclusive measure in a biological context. Cell conditions, such as differentiation or infection, are known to yield significant changes in the refractive index. Furthermore, the refractive index of biological tissue determines the way it interacts with light. Besides the biological relevance of this interaction in the retina, a lot of methods used in biology, including microscopy, rely on light-tissue or light-cell interactions. Hence, determining the refractive index of cells using DHM is valuable in many biological applications. This chapter covers the main topics which are important for the implementation of DHM: setup, sample preparation and analysis. First, the optical setup is described in detail including notes and suggestions for the implementation. Following that, a protocol for the sample and measurement preparation is explained. In the analysis section, an algorithm for the determination of the quantitative phase map is described. Subsequently, all intermediate steps for the calculation of the refractive index of suspended cells are presented, exploiting their spherical shape. In the last section, a discussion of possible extensions to the setup, further measurement configurations and additional analysis methods are given. Throughout this chapter, we describe a simple, robust, and thus easily reproducible implementation of DHM. The different possibilities for extensions show the diverse fields of application for this technique., Comment: 15 pages, 3 figures

Measuring the average refractive index (RI) of spherical objects, such as suspended cells, in quantitative phase imaging (QPI) requires a decoupling of RI and size from the QPI data. This has been commonly achieved by determining the object's radius with geometrical approaches, neglecting light-scattering. Here, we present a novel QPI fitting algorithm that reliably uncouples the RI using Mie theory and a semi-analytical, corrected Rytov approach. We assess the range of validity of this algorithm in silico and experimentally investigate various objects (oil and protein droplets, microgel beads, cells) and noise conditions. In addition, we provide important practical cues for future studies in cell biology., Comment: 14 pages, 10 figures, 1 table

Optical diffraction tomography (ODT) is a tomographic technique that can be used to measure the three-dimensional (3D) refractive index distribution within living cells without the requirement of any marker. In principle, ODT can be regarded as a generalization of optical projection tomography which is equivalent to computerized tomography (CT). Both optical tomographic techniques require projection-phase images of cells measured at multiple angles. However, the reconstruction of the 3D refractive index distribution post-measurement differs for the two techniques. It is known that ODT yields better results than projection tomography, because it takes into account diffraction of the imaging light due to the refractive index structure of the sample. Here, we apply ODT to biological cells in a microfluidic chip which combines optical trapping and microfluidic flow to achieve an optofluidic single-cell rotation. In particular, we address the problem that arises when the trapped cell is not rotating about an axis perpendicular to the imaging plane, but instead about an arbitrarily tilted axis. In this paper we show that the 3D reconstruction can be improved by taking into account such a tilted rotational axis in the reconstruction process., Comment: 7 pages, 3 figures

Cells alter the path of light, a fact that leads to well-known aberrations in single cell or tissue imaging. Optical diffraction tomography (ODT) measures the biophysical property that causes these aberrations, the refractive index (RI). ODT is complementary to fluorescence imaging and does not require any markers. The present study introduces RI and fluorescence tomography with optofluidic rotation (RAFTOR) of suspended cells, quantifying the intracellular RI distribution and colocalizing it with fluorescence in 3D. The technique is validated with cell phantoms and used to confirm a lower nuclear RI for HL60 cells. Furthermore, the nuclear inversion of adult mouse photoreceptor cells is observed in the RI distribution. The applications shown confirm predictions of previous studies and illustrate the potential of RAFTOR to improve our understanding of cells and tissues., Comment: 15 pages, 5 figures

Tomography is the three-dimensional reconstruction of an object from images taken at different angles. The term classical tomography is used, when the imaging beam travels in straight lines through the object. This assumption is valid for light with short wavelengths, for example in x-ray tomography. For classical tomography, a commonly used reconstruction method is the filtered back-projection algorithm which yields fast and stable object reconstructions. In the context of single-cell imaging, the back-projection algorithm has been used to investigate the cell structure or to quantify the refractive index distribution within single cells using light from the visible spectrum. Nevertheless, these approaches, commonly summarized as optical projection tomography, do not take into account diffraction. Diffraction tomography with the Rytov approximation resolves this issue. The explicit incorporation of the wave nature of light results in an enhanced reconstruction of the object's refractive index distribution. Here, we present a full literature review of diffraction tomography. We derive the theory starting from the wave equation and discuss its validity with the focus on applications for refractive index tomography. Furthermore, we derive the back-propagation algorithm, the diffraction-tomographic pendant to the back-projection algorithm, and describe its implementation in three dimensions. Finally, we showcase the application of the back-propagation algorithm to computer-generated scattering data. This review unifies the different notations in literature and gives a detailed description of the back-propagation algorithm, serving as a reliable basis for future work in the field of diffraction tomography., Comment: 59 pages, 10 figures, 4 tables