@inproceedings{21629, abstract = {We measure the second-order coherence function g(2) for X-ray-driven light emission (scintillation), observing that it is bunched (g(2) > 1), and can achieve extreme bunching values (g(2)~97) in perovskite nano-crystals.}, author = {Katznelson, Shaul and Tziperman, Offek and Bucher, Tomer and Abudi, Tom Lenkiewicz and Schuetz, Roman and Be'er, Orr and Levy, Shai and Bekenstein, Yehonadav and Roques-Carmes, Charles and Kaminer, Ido}, booktitle = {Conference on Lasers and Electro-Optics}, location = {San Jose, CA, United States}, publisher = {Optica Publishing Group}, title = {{X-ray-driven photon bunching}}, doi = {10.1364/cleo_si.2023.sm1h.6}, year = {2023}, } @inproceedings{21595, abstract = {We present a method for x-ray spectroscopy, combining nanophotonic scintillator inverse design with an image reconstruction algorithm. We demonstrate our pipeline on 3-energy x-ray spectroscopy, achieving 8% reconstruction error under 1% Gaussian noise}, author = {Li, William F. and Roques-Carmes, Charles and Lin, Zin and Johnson, Steven G. and Soljačić, Marin}, booktitle = {Conference on Lasers and Electro-Optics}, location = {San Jose, CA, United States}, publisher = {Optica Publishing Group}, title = {{X-ray spectroscopy with end-to-end optimized nanophotonic scintillators}}, doi = {10.1364/cleo_fs.2023.fw4c.4}, year = {2023}, } @article{21810, abstract = {The next-generation semiconductors and devices, such as halide perovskites and flexible electronics, are extremely sensitive to water, thus demanding highly effective protection that not only seals out water in all forms (vapor, droplet, and ice), but simultaneously provides mechanical flexibility, durability, transparency, and self-cleaning. Although various solid-state encapsulation methods have been developed, no strategy is available that can fully meet all the above requirements. Here, we report a bioinspired liquid-based encapsulation strategy that offers protection from water without sacrificing the operational properties of the encapsulated materials. Using halide perovskite as a model system, we show that damage to the perovskite from exposure to water is drastically reduced when it is coated by a polymer matrix with infused hydrophobic oil. With a combination of experimental and simulation studies, we elucidated the fundamental transport mechanisms of ultralow water transmission rate that stem from the ability of the infused liquid to fill-in and reduce defects in the coating layer, thus eliminating the low-energy diffusion pathways, and to cause water molecules to diffuse as clusters, which act together as an excellent water permeation barrier. Importantly, the presence of the liquid, as the central component in this encapsulation method provides a unique possibility of reversing the water transport direction; therefore, the lifetime of enclosed water-sensitive materials could be significantly extended via replenishing the hydrophobic oils regularly. We show that the liquid encapsulation platform presented here has high potential in providing not only water protection of the functional device but also flexibility, optical transparency, and self-healing of the coating layer, which are critical for a variety of applications, such as in perovskite solar cells and bioelectronics.}, author = {Lemaire, Baptiste and Yu, Yanhao and Molinari, Nicola and Wu, Haichao and Goodwin, Zachary A. H. and Stricker, Friedrich J and Kozinsky, Boris and Aizenberg, Joanna}, issn = {1091-6490}, journal = {Proceedings of the National Academy of Sciences}, keywords = {water permeability, photoelectronic materials, device encapsulation, liquid-infused polymers}, number = {34}, publisher = {National Academy of Sciences}, title = {{Flexible fluid-based encapsulation platform for water-sensitive materials}}, doi = {10.1073/pnas.2308804120}, volume = {120}, year = {2023}, } @article{21813, abstract = {Aligned liquid crystal polymers are materials of interest for electronic, optic, biological and soft robotic applications. The manufacturing and processing of these materials have been widely explored with mechanical alignment establishing itself as a preferred method due to its ease of use and widespread applicability. However, the fundamental chemistry behind the required two‐step polymerization for mechanical alignment has limitations in both fabrication and substrate compatibility. In this work we introduce a new protection‐deprotection approach utilizing a two‐stage Diels–Alder cyclopentadiene‐maleimide step‐growth polymerization to enable mild yet efficient, fast, controlled, reproducible and user‐friendly polymerizations, broadening the scope of liquid crystal systems. Thorough characterization of the films by DSC, DMA, POM and WAXD show the successful synthesis of a uniaxially aligned liquid crystal network with thermomechanical actuation abilities.}, author = {Guillen Campos, Jesus and Stricker, Friedrich J and Clark, Kyle D. and Park, Minwook and Bailey, Sophia J. and Kuenstler, Alexa S. and Hayward, Ryan C. and Read de Alaniz, Javier}, issn = {1521-3773}, journal = {Angewandte Chemie International Edition}, number = {1}, publisher = {Wiley}, title = {{Controlled Diels–Alder “Click” strategy to access mechanically aligned main‐chain liquid crystal networks}}, doi = {10.1002/anie.202214339}, volume = {62}, year = {2023}, } @article{21807, abstract = {Multifaceted material responses upon exposure to stimuli are key for developing life-like materials. Developing such synthetic systems, though not trivial, typically relies on orthogonal stimuli to enable control of molecular systems that enable multi-responsive behavior. Access to complex tunable reaction mechanisms with diverse energy landscapes offers an alternative strategy for controlling out-of-equilibrium processes without requiring orthogonal stimuli for each responsive unit. Donor-acceptor Stenhouse adducts (DASAs) are a class of photoswitches that have complex, tunable, and environmentally sensitive reaction pathways. We present the control of donor-acceptor Stenhouse adduct equilibrium and photoswitching kinetics through changes in the polarity of their environment. Polarity and light can be used to selectively control the pathway outcomes of three DASA derivatives where the orthogonal response comes from changes in the energy landscape and is not driven by their orthogonal response to the given stimuli. This work paves the way to designing multi-responsive and self-regulating life-like materials.}, author = {Stricker, Friedrich J and Peterson, Julie and Sandlass, Sara K. and de Tagyos, Aurora and Sroda, Miranda and Seshadri, Serena and Gordon, Michael J. and Read de Alaniz, Javier}, issn = {2451-9294}, journal = {Chem}, number = {7}, pages = {1994--2005}, publisher = {Elsevier}, title = {{Selective control of donor-acceptor Stenhouse adduct populations with non-selective stimuli}}, doi = {10.1016/j.chempr.2023.05.011}, volume = {9}, year = {2023}, } @article{21818, abstract = {Surface-aligned liquid-crystal networks (LCNs) offer a solution for developing functional materials capable of performing a range of tasks, including actuation, shape memory, and surfaces patterning. Here we show that Diels–Alder cycloaddition can be used to prepare the backbone of planar aligned LCNs under mild ambient conditions without the addition of additives or UV irradiation. The mechanical properties of the networks have robust viscoelastic modulus and stiffness with a reversible local free volume change upon physical aging. This study shows new opportunities to design surface-aligned LCNs based on additive free step-growth Diels–Alder polymerization and enables the potential to incorporate a wider range of photochromic materials into LCNs.}, author = {Park, Minwook and Stricker, Friedrich J and Campos, Jesus Guillen and Clark, Kyle D. and Lee, Jaejun and Kwon, Younghoon and Valentine, Megan T. and Read de Alaniz, Javier}, issn = {2161-1653}, journal = {ACS Macro Letters}, number = {1}, pages = {33--39}, publisher = {American Chemical Society}, title = {{Design of surface-aligned main-chain liquid-crystal networks prepared under ambient, light-free conditions using the Diels–Alder cycloaddition}}, doi = {10.1021/acsmacrolett.2c00616}, volume = {12}, year = {2023}, } @article{21821, abstract = {Molecular photoswitches provide a means for imparting synthetic structures with intrinsically logical and highly tunable photoresponsive properties. One variety of organic photoswitches known as Donor-Acceptor Stenhouse Adducts, or DASAs, are promising candidates for next generation light responsive materials because of their unique ability to stabilize three photochemically distinct isomeric states in solution, while their counterparts are strictly limited to binary state behavior. In this work, we show how polymethacrylate host matrices shift the energetic landscape of DASA relative to solution, prohibiting accumulation of an intermediate third isomeric state by decelerating critical steps in the photoswitching mechanism. Specifically, we employ a dual-wavelength, phase locked detection scheme to probe thermal isomerizations in the switching process that occur at fast (~ms) time scales that are inaccessible by standard UV–Vis spectroscopic techniques. The results of this study provide valuable insight into the mechanism of multistate DASA reactivity and establish the foundation necessary to guide future efforts in offsetting kinetic matrix effects to enable dynamic, three state photoswitching in polymeric hosts. }, author = {Sandlass, Sara and Stricker, Friedrich J and Fragoso, Daniel and de Alaniz, Javier Read and Gordon, Michael J.}, issn = {1873-2666}, journal = {Journal of Photochemistry and Photobiology A: Chemistry}, publisher = {Elsevier}, title = {{Effect of polymer host matrix on multi-stage isomerization kinetics of DASA photochromes}}, doi = {10.1016/j.jphotochem.2023.114964}, volume = {444}, year = {2023}, } @inproceedings{14771, abstract = {Pruning—that is, setting a significant subset of the parameters of a neural network to zero—is one of the most popular methods of model compression. Yet, several recent works have raised the issue that pruning may induce or exacerbate bias in the output of the compressed model. Despite existing evidence for this phenomenon, the relationship between neural network pruning and induced bias is not well-understood. In this work, we systematically investigate and characterize this phenomenon in Convolutional Neural Networks for computer vision. First, we show that it is in fact possible to obtain highly-sparse models, e.g. with less than 10% remaining weights, which do not decrease in accuracy nor substantially increase in bias when compared to dense models. At the same time, we also find that, at higher sparsities, pruned models exhibit higher uncertainty in their outputs, as well as increased correlations, which we directly link to increased bias. We propose easy-to-use criteria which, based only on the uncompressed model, establish whether bias will increase with pruning, and identify the samples most susceptible to biased predictions post-compression. Our code can be found at https://github.com/IST-DASLab/pruned-vision-model-bias.}, author = {Iofinova, Eugenia B and Peste, Elena-Alexandra and Alistarh, Dan-Adrian}, booktitle = {2023 IEEE/CVF Conference on Computer Vision and Pattern Recognition}, issn = {2575-7075}, location = {Vancouver, BC, Canada}, pages = {24364--24373}, publisher = {IEEE}, title = {{Bias in pruned vision models: In-depth analysis and countermeasures}}, doi = {10.1109/cvpr52729.2023.02334}, year = {2023}, } @phdthesis{13286, abstract = {Semiconductor-superconductor hybrid systems are the harbour of many intriguing mesoscopic phenomena. This material combination leads to spatial variations of the superconducting properties, which gives rise to Andreev bound states (ABSs). Some of these states might exhibit remarkable properties that render them highly desirable for topological quantum computing. The most prominent and hunted of such states are Majorana zero modes (MZMs), quasiparticles equals to their own quasiparticles that they follow non-abelian statistics. In this thesis, we first introduce the general framework of such hybrid systems and, then, we unveil a series of mesoscopic phenomena that we discovered. Firstly, we show tunneling spectroscopy experiments on full-shell nanowires (NWs) showing that unwanted quantum-dot states coupled to superconductors (Yu-Shiba-Rusinov states) can mimic MZMs signatures. Then, we introduce a novel protocol which allowed the integration of tunneling spectroscopy with Coulomb spectroscopy within the same device. Employing this approach on both full-shell NWs and partial-shell NWs, we demonstrated that longitudinally confined states reveal charge transport phenomenology similar to the one expected for MZMs. These findings shed light on the intricate interplay between superconductivity and quantum confinement, which brought us to explore another material platform, i.e. a two-dimensional Germanium hole gas. After developing a robust way to induce superconductivity in such system, we showed how to engineer the proximity effect and we revealed a superconducting hard gap. Finally, we created a superconducting radio frequency driven ideal diode and a generator of non-sinusoidal current-phase relations. Our results open the path for the exploration of protected superconducting qubits and more complex hybrid devices in planar Germanium, like Kitaev chains and hybrid qubit devices.}, author = {Valentini, Marco}, issn = {2663-337X}, pages = {184}, publisher = {Institute of Science and Technology Austria}, title = {{Mesoscopic phenomena in hybrid semiconductor-superconductor nanodevices : From full-shell nanowires to two-dimensional hole gas in germanium}}, doi = {10.15479/at:ista:13286}, year = {2023}, } @article{14517, abstract = {State-of-the-art transmon qubits rely on large capacitors, which systematically improve their coherence due to reduced surface-loss participation. However, this approach increases both the footprint and the parasitic cross-coupling and is ultimately limited by radiation losses—a potential roadblock for scaling up quantum processors to millions of qubits. In this work we present transmon qubits with sizes as low as 36 × 39 µm2 with 100-nm-wide vacuum-gap capacitors that are micromachined from commercial silicon-on-insulator wafers and shadow evaporated with aluminum. We achieve a vacuum participation ratio up to 99.6% in an in-plane design that is compatible with standard coplanar circuits. Qubit relaxationtime measurements for small gaps with high zero-point electric field variance of up to 22 V/m reveal a double exponential decay indicating comparably strong qubit interaction with long-lived two-level systems. The exceptionally high selectivity of up to 20 dB to the superconductor-vacuum interface allows us to precisely back out the sub-single-photon dielectric loss tangent of aluminum oxide previously exposed to ambient conditions. In terms of future scaling potential, we achieve a ratio of qubit quality factor to a footprint area equal to 20 µm−2, which is comparable with the highest T1 devices relying on larger geometries, a value that could improve substantially for lower surface-loss superconductors. }, author = {Zemlicka, Martin and Redchenko, Elena and Peruzzo, Matilda and Hassani, Farid and Trioni, Andrea and Barzanjeh, Shabir and Fink, Johannes M}, issn = {2331-7019}, journal = {Physical Review Applied}, number = {4}, publisher = {American Physical Society}, title = {{Compact vacuum-gap transmon qubits: Selective and sensitive probes for superconductor surface losses}}, doi = {10.1103/PhysRevApplied.20.044054}, volume = {20}, year = {2023}, } @unpublished{13312, abstract = {Superconductor/semiconductor hybrid devices have attracted increasing interest in the past years. Superconducting electronics aims to complement semiconductor technology, while hybrid architectures are at the forefront of new ideas such as topological superconductivity and protected qubits. In this work, we engineer the induced superconductivity in two-dimensional germanium hole gas by varying the distance between the quantum well and the aluminum. We demonstrate a hard superconducting gap and realize an electrically and flux tunable superconducting diode using a superconducting quantum interference device (SQUID). This allows to tune the current phase relation (CPR), to a regime where single Cooper pair tunneling is suppressed, creating a $ \sin \left( 2 \varphi \right)$ CPR. Shapiro experiments complement this interpretation and the microwave drive allows to create a diode with $ \approx 100 \%$ efficiency. The reported results open up the path towards monolithic integration of spin qubit devices, microwave resonators and (protected) superconducting qubits on a silicon technology compatible platform.}, author = {Valentini, Marco and Sagi, Oliver and Baghumyan, Levon and Gijsel, Thijs de and Jung, Jason and Calcaterra, Stefano and Ballabio, Andrea and Servin, Juan Aguilera and Aggarwal, Kushagra and Janik, Marian and Adletzberger, Thomas and Souto, Rubén Seoane and Leijnse, Martin and Danon, Jeroen and Schrade, Constantin and Bakkers, Erik and Chrastina, Daniel and Isella, Giovanni and Katsaros, Georgios}, booktitle = {arXiv}, keywords = {Mesoscale and Nanoscale Physics}, title = {{Radio frequency driven superconducting diode and parity conserving Cooper pair transport in a two-dimensional germanium hole gas}}, doi = {10.48550/arXiv.2306.07109}, year = {2023}, } @misc{13124, abstract = {This dataset comprises all data shown in the figures of the submitted article "Tunable directional photon scattering from a pair of superconducting qubits" at arXiv:2205.03293. Additional raw data are available from the corresponding author on reasonable request.}, author = {Redchenko, Elena and Poshakinskiy, Alexander and Sett, Riya and Zemlicka, Martin and Poddubny, Alexander and Fink, Johannes M}, publisher = {Zenodo}, title = {{Tunable directional photon scattering from a pair of superconducting qubits}}, doi = {10.5281/ZENODO.7858567}, year = {2023}, } @article{13314, abstract = {The emergence of large-scale order in self-organized systems relies on local interactions between individual components. During bacterial cell division, FtsZ—a prokaryotic homologue of the eukaryotic protein tubulin—polymerizes into treadmilling filaments that further organize into a cytoskeletal ring. In vitro, FtsZ filaments can form dynamic chiral assemblies. However, how the active and passive properties of individual filaments relate to these large-scale self-organized structures remains poorly understood. Here we connect single-filament properties with the mesoscopic scale by combining minimal active matter simulations and biochemical reconstitution experiments. We show that the density and flexibility of active chiral filaments define their global order. At intermediate densities, curved, flexible filaments organize into chiral rings and polar bands. An effectively nematic organization dominates for high densities and for straight, mutant filaments with increased rigidity. Our predicted phase diagram quantitatively captures these features, demonstrating how the flexibility, density and chirality of the active filaments affect their collective behaviour. Our findings shed light on the fundamental properties of active chiral matter and explain how treadmilling FtsZ filaments organize during bacterial cell division.}, author = {Dunajova, Zuzana and Prats Mateu, Batirtze and Radler, Philipp and Lim, Keesiang and Brandis, Dörte and Velicky, Philipp and Danzl, Johann G and Wong, Richard W. and Elgeti, Jens and Hannezo, Edouard B and Loose, Martin}, issn = {1745-2481}, journal = {Nature Physics}, pages = {1916--1926}, publisher = {Springer Nature}, title = {{Chiral and nematic phases of flexible active filaments}}, doi = {10.1038/s41567-023-02218-w}, volume = {19}, year = {2023}, } @article{14759, abstract = {Proper operation of electro-optic I/Q modulators relies on precise adjustment and control of the relative phase biases between the modulator’s internal interferometer arms. We present an all-analog phase bias locking scheme where error signals are obtained from the beat between the optical carrier and optical tones generated by an auxiliary 2 MHz 𝑅𝐹 tone to lock the phases of all three involved interferometers for operation up to 10 GHz. With the developed method, we demonstrate an I/Q modulator in carrier-suppressed single-sideband mode, where the suppressed carrier and sideband are locked at optical power levels <−27dB relative to the transmitted sideband. We describe a simple analytical model for calculating the error signals and detail the implementation of the electronic circuitry for the implementation of the method.}, author = {Wald, Sebastian and Diorico, Fritz R and Hosten, Onur}, issn = {2155-3165}, journal = {Applied Optics}, keywords = {Atomic and Molecular Physics, and Optics, Engineering (miscellaneous), Electrical and Electronic Engineering}, number = {1}, pages = {1--7}, publisher = {Optica Publishing Group}, title = {{Analog stabilization of an electro-optic I/Q modulator with an auxiliary modulation tone}}, doi = {10.1364/ao.474118}, volume = {62}, year = {2023}, } @inproceedings{14459, abstract = {Autoencoders are a popular model in many branches of machine learning and lossy data compression. However, their fundamental limits, the performance of gradient methods and the features learnt during optimization remain poorly understood, even in the two-layer setting. In fact, earlier work has considered either linear autoencoders or specific training regimes (leading to vanishing or diverging compression rates). Our paper addresses this gap by focusing on non-linear two-layer autoencoders trained in the challenging proportional regime in which the input dimension scales linearly with the size of the representation. Our results characterize the minimizers of the population risk, and show that such minimizers are achieved by gradient methods; their structure is also unveiled, thus leading to a concise description of the features obtained via training. For the special case of a sign activation function, our analysis establishes the fundamental limits for the lossy compression of Gaussian sources via (shallow) autoencoders. Finally, while the results are proved for Gaussian data, numerical simulations on standard datasets display the universality of the theoretical predictions.}, author = {Shevchenko, Aleksandr and Kögler, Kevin and Hassani, Hamed and Mondelli, Marco}, booktitle = {Proceedings of the 40th International Conference on Machine Learning}, issn = {2640-3498}, location = {Honolulu, Hawaii, HI, United States}, pages = {31151--31209}, publisher = {ML Research Press}, title = {{Fundamental limits of two-layer autoencoders, and achieving them with gradient methods}}, volume = {202}, year = {2023}, } @phdthesis{12897, abstract = {Inverse design problems in fabrication-aware shape optimization are typically solved on discrete representations such as polygonal meshes. This thesis argues that there are benefits to treating these problems in the same domain as human designers, namely, the parametric one. One reason is that discretizing a parametric model usually removes the capability of making further manual changes to the design, because the human intent is captured by the shape parameters. Beyond this, knowledge about a design problem can sometimes reveal a structure that is present in a smooth representation, but is fundamentally altered by discretizing. In this case, working in the parametric domain may even simplify the optimization task. We present two lines of research that explore both of these aspects of fabrication-aware shape optimization on parametric representations. The first project studies the design of plane elastic curves and Kirchhoff rods, which are common mathematical models for describing the deformation of thin elastic rods such as beams, ribbons, cables, and hair. Our main contribution is a characterization of all curved shapes that can be attained by bending and twisting elastic rods having a stiffness that is allowed to vary across the length. Elements like these can be manufactured using digital fabrication devices such as 3d printers and digital cutters, and have applications in free-form architecture and soft robotics. We show that the family of curved shapes that can be produced this way admits geometric description that is concise and computationally convenient. In the case of plane curves, the geometric description is intuitive enough to allow a designer to determine whether a curved shape is physically achievable by visual inspection alone. We also present shape optimization algorithms that convert a user-defined curve in the plane or in three dimensions into the geometry of an elastic rod that will naturally deform to follow this curve when its endpoints are attached to a support structure. Implemented in an interactive software design tool, the rod geometry is generated in real time as the user edits a curve and enables fast prototyping. The second project tackles the problem of general-purpose shape optimization on CAD models using a novel variant of the extended finite element method (XFEM). Our goal is the decoupling between the simulation mesh and the CAD model, so no geometry-dependent meshing or remeshing needs to be performed when the CAD parameters change during optimization. This is achieved by discretizing the embedding space of the CAD model, and using a new high-accuracy numerical integration method to enable XFEM on free-form elements bounded by the parametric surface patches of the model. Our simulation is differentiable from the CAD parameters to the simulation output, which enables us to use off-the-shelf gradient-based optimization procedures. The result is a method that fits seamlessly into the CAD workflow because it works on the same representation as the designer, enabling the alternation of manual editing and fabrication-aware optimization at will.}, author = {Hafner, Christian}, isbn = {978-3-99078-031-2}, issn = {2663-337X}, pages = {180}, publisher = {Institute of Science and Technology Austria}, title = {{Inverse shape design with parametric representations: Kirchhoff Rods and parametric surface models}}, doi = {10.15479/at:ista:12897}, year = {2023}, } @article{13188, abstract = {The Kirchhoff rod model describes the bending and twisting of slender elastic rods in three dimensions, and has been widely studied to enable the prediction of how a rod will deform, given its geometry and boundary conditions. In this work, we study a number of inverse problems with the goal of computing the geometry of a straight rod that will automatically deform to match a curved target shape after attaching its endpoints to a support structure. Our solution lets us finely control the static equilibrium state of a rod by varying the cross-sectional profiles along its length. We also show that the set of physically realizable equilibrium states admits a concise geometric description in terms of linear line complexes, which leads to very efficient computational design algorithms. Implemented in an interactive software tool, they allow us to convert three-dimensional hand-drawn spline curves to elastic rods, and give feedback about the feasibility and practicality of a design in real time. We demonstrate the efficacy of our method by designing and manufacturing several physical prototypes with applications to interior design and soft robotics.}, author = {Hafner, Christian and Bickel, Bernd}, issn = {1557-7368}, journal = {ACM Transactions on Graphics}, keywords = {Computer Graphics, Computational Design, Computational Geometry, Shape Modeling}, number = {5}, publisher = {Association for Computing Machinery}, title = {{The design space of Kirchhoff rods}}, doi = {10.1145/3606033}, volume = {42}, year = {2023}, } @phdthesis{12491, abstract = {The extracellular matrix (ECM) is a hydrated and complex three-dimensional network consisting of proteins, polysaccharides, and water. It provides structural scaffolding for the cells embedded within it and is essential in regulating numerous physiological processes, including cell migration and proliferation, wound healing, and stem cell fate. Despite extensive study, detailed structural knowledge of ECM components in physiologically relevant conditions is still rudimentary. This is due to methodological limitations in specimen preparation protocols which are incompatible with keeping large samples, such as the ECM, in their native state for subsequent imaging. Conventional electron microscopy (EM) techniques rely on fixation, dehydration, contrasting, and sectioning. This results in the alteration of a highly hydrated environment and the potential introduction of artifacts. Other structural biology techniques, such as nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography, allow high-resolution analysis of protein structures but only work on homogenous and purified samples, hence lacking contextual information. Currently, no approach exists for the ultrastructural and structural study of extracellular components under native conditions in a physiological, 3D environment. In this thesis, I have developed a workflow that allows for the ultrastructural analysis of the ECM in near-native conditions at molecular resolution. The developments I introduced include implementing a novel specimen preparation workflow for cell-derived matrices (CDMs) to render them compatible with ion-beam milling and subsequent high-resolution cryo-electron tomography (ET). To this end, I have established protocols to generate CDMs grown over several weeks on EM grids that are compatible with downstream cryo-EM sample preparation and imaging techniques. Characterization of these ECMs confirmed that they contain essential ECM components such as collagen I, collagen VI, and fibronectin I in high abundance and hence represent a bona fide biologically-relevant sample. I successfully optimized vitrification of these specimens by testing various vitrification techniques and cryoprotectants. In order to obtain high-resolution molecular insights into the ultrastructure and organization of CDMs, I established cryo-focused ion beam scanning electron microscopy (FIBSEM) on these challenging and complex specimens. I explored different approaches for the creation of thin cryo-lamellae by FIB milling and succeeded in optimizing the cryo-lift-out technique, resulting in high-quality lamellae of approximately 200 nm thickness. High-resolution Cryo-ET of these lamellae revealed for the first time the architecture of native CDM in the context of matrix-secreting cells. This allowed for the in situ visualization of fibrillar matrix proteins such as collagen, laying the foundation for future structural and ultrastructural characterization of these proteins in their near-native environment. In summary, in this thesis, I present a novel workflow that combines state-of-the-art cryo-EM specimen preparation and imaging technologies to permit characterization of the ECM, an important tissue component in higher organisms. This innovative and highly versatile workflow will enable addressing far-reaching questions on ECM architecture, composition, and reciprocal ECM-cell interactions.}, author = {Zens, Bettina}, isbn = {978-3-99078-027-5}, issn = {2663-337X}, keywords = {cryo-EM, cryo-ET, FIB milling, method development, FIBSEM, extracellular matrix, ECM, cell-derived matrices, CDMs, cell culture, high pressure freezing, HPF, structural biology, tomography, collagen}, pages = {187}, publisher = {Institute of Science and Technology Austria}, title = {{Ultrastructural characterization of natively preserved extracellular matrix by cryo-electron tomography}}, doi = {10.15479/at:ista:12491}, year = {2023}, } @phdthesis{13984, abstract = {Social insects fight disease using their individual immune systems and the cooperative sanitary behaviors of colony members. These social defenses are well explored against externally-infecting pathogens, but little is known about defense strategies against internally-infecting pathogens, such as viruses. Viruses are ubiquitous and in the last decades it has become evident that also many ant species harbor viruses. We present one of the first studies addressing transmission dynamics and collective disease defenses against viruses in ants on a mechanistic level. I successfully established an experimental ant host – viral pathogen system as a model for the defense strategies used by social insects against internal pathogen infections, as outlined in the third chapter. In particular, we studied how garden ants (Lasius neglectus) defend themselves and their colonies against the generalist insect virus CrPV (cricket paralysis virus). We chose microinjections of virus directly into the ants’ hemolymph because it allowed us to use a defined exposure dose. Here we show that this is a good model system, as the virus is replicating and thus infecting the host. The ants mount a clear individual immune response against the viral infection, which is characterized by a specific siRNA pattern, namely siRNAs mapping against the viral genome with a peak of 21 and 22 bp long fragments. The onset of this immune response is consistent with the timeline of viral replication that starts already within two days post injection. The disease manifests in decreased survival over a course of two to three weeks. Regarding group living, we find that infected ants show a strong individual immune response, but that their course of disease is little affected by nestmate presence, as described in chapter four. Hence, we do not find social immunity in the context of viral infections in ants. Nestmates, however, can contract the virus. Using Drosophila S2R+ cells in culture, we showed that 94 % of the nestmates contract active virus within four days of social contact to an infected individual. Virus is transmitted in low doses, thus not causing disease transmission within the colony. While virus can be transmitted during short direct contacts, we also assume transmission from deceased ants and show that the nestmates’ immune system gets activated after contracting a low viral dose. We find considerable potential for indirect transmission via the nest space. Virus is shed to the nest, where it stays viable for one week and is also picked up by other ants. Apart from that, we want to underline the potential of ant poison as antiviral agent. We determined that ant poison successfully inactivates CrPV in vitro. However, we found no evidence for effective poison use to sanitize the nest space. On the other hand, local application of ant poison by oral poison uptake, which is part of the ants prophylactic behavioral repertoire, probably contributes to keeping the gut of each individual sanitized. We hypothesize that oral poison uptake might be the reason why we did not find viable virus in the trophallactic fluid. The fifth chapter encompasses preliminary data on potential social immunization. However, our experiments do not confirm an actual survival benefit for the nestmates upon pathogen challenge under the given experimental settings. Nevertheless, we do not want to rule out the possibility for nestmate immunization, but rather emphasize that considering different experimental timelines and viral doses would provide a multitude of options for follow-up experiments. In conclusion, we find that prophylactic individual behaviors, such as oral poison uptake, might play a role in preventing viral disease transmission. Compared to colony defense against external pathogens, internal pathogen infections require a stronger component of individual physiological immunity than behavioral social immunity, yet could still lead to collective protection.}, author = {Franschitz, Anna}, isbn = {978-3-99078-034-3}, issn = {2663-337X}, pages = {89}, publisher = {Institute of Science and Technology Austria}, title = {{Individual and social immunity against viral infections in ants}}, doi = {10.15479/at:ista:13984}, year = {2023}, } @phdthesis{12964, abstract = {Pattern formation is of great importance for its contribution across different biological behaviours. During developmental processes for example, patterns of chemical gradients are established to determine cell fate and complex tissue patterns emerge to define structures such as limbs and vascular networks. Patterns are also seen in collectively migrating groups, for instance traveling waves of density emerging in moving animal flocks as well as collectively migrating cells and tissues. To what extent these biological patterns arise spontaneously through the local interaction of individual constituents or are dictated by higher level instructions is still an open question however there is evidence for the involvement of both types of process. Where patterns arise spontaneously there is a long standing interest in how far the interplay of mechanics, e.g. force generation and deformation, and chemistry, e.g. gene regulation and signaling, contributes to the behaviour. This is because many systems are able to both chemically regulate mechanical force production and chemically sense mechanical deformation, forming mechano-chemical feedback loops which can potentially become unstable towards spatio and/or temporal patterning. We work with experimental collaborators to investigate the possibility that this type of interaction drives pattern formation in biological systems at different scales. We focus first on tissue-level ERK-density waves observed during the wound healing response across different systems where many previous studies have proposed that patterns depend on polarized cell migration and arise from a mechanical flocking-like mechanism. By combining theory with mechanical and optogenetic perturbation experiments on in vitro monolayers we instead find evidence for mechanochemical pattern formation involving only scalar bilateral feedbacks between ERK signaling and cell contraction. We perform further modeling and experiment to study how this instability couples with polar cell migration in order to produce a robust and efficient wound healing response. In a following chapter we implement ERK-density coupling and cell migration in a 2D active vertex model to investigate the interaction of ERK-density patterning with different tissue rheologies and find that the spatio-temporal dynamics are able to both locally and globally fluidize a tissue across the solid-fluid glass transition. In a last chapter we move towards lower spatial scales in the context of subcellular patterning of the cell cytoskeleton where we investigate the transition between phases of spatially homogeneous temporal oscillations and chaotic spatio-temporal patterning in the dynamics of myosin and ROCK activities (a motor component of the actomyosin cytoskeleton and its activator). Experimental evidence supports an intrinsic chemical oscillator which we encode in a reaction model and couple to a contractile active gel description of the cell cortex. The model exhibits phases of chemical oscillations and contractile spatial patterning which reproduce many features of the dynamics seen in Drosophila oocyte epithelia in vivo. However, additional pharmacological perturbations to inhibit myosin contractility leaves the role of contractile instability unclear. We discuss alternative hypotheses and investigate the possibility of reaction-diffusion instability.}, author = {Boocock, Daniel R}, isbn = {978-3-99078-032-9}, issn = {2663-337X}, pages = {146}, publisher = {Institute of Science and Technology Austria}, title = {{Mechanochemical pattern formation across biological scales}}, doi = {10.15479/at:ista:12964}, year = {2023}, }