Search

Single-molecule localization microscopy generates point clouds corresponding to fluorophore localizations. Spatial cluster identification and analysis of these point clouds are crucial for extracting insights about molecular organization. However, this task becomes challenging in the presence of localization noise, high point density, or complex biological structures. Here, we introduce MIRO (Multimodal Integration through Relational Optimization), an algorithm that uses recurrent graph neural networks to transform the point clouds in order to improve clustering efficiency when applying conventional clustering techniques. We show that MIRO supports simultaneous processing of clusters of different shapes and at multiple scales, demonstrating improved performance across varied datasets. Our comprehensive evaluation demonstrates MIRO's transformative potential for single-molecule localization applications, showcasing its capability to revolutionize cluster analysis and provide accurate, reliable details of molecular architecture. In addition, MIRO's robust clustering capabilities hold promise for applications in various fields such as neuroscience, for the analysis of neural connectivity patterns, and environmental science, for studying spatial distributions of ecological data.

The miniaturization of mechanical machines is critical for advancing nanotechnology and reducing device footprints. Traditional efforts to downsize gears and micromotors have faced limitations at around 0.1 mm for over thirty years due to the complexities of constructing drives and coupling systems at such scales. Here, we present an alternative approach utilizing optical metasurfaces to locally drive microscopic machines, which can then be fabricated using standard lithography techniques and seamlessly integrated on the chip, achieving sizes down to tens of micrometers with movements precise to the sub-micrometer scale. As a proof of principle, we demonstrate the construction of microscopic gear trains powered by a single driving gear with a metasurface activated by a plane light wave. Additionally, we develop a versatile pinion and rack micromachine capable of transducing rotational motion, performing periodic motion, and controlling microscopic mirrors for light deflection. Our on-chip fabrication process allows for straightforward parallelization and integration. Using light as a widely available and easily controllable energy source, these miniaturized metamachines offer precise control and movement, unlocking new possibilities for micro- and nanoscale systems.

Diffusion models have emerged as a prominent technique in generative modeling with neural networks, making their mark in tasks like text-to-image translation and super-resolution. In this tutorial, we provide a comprehensive guide to build denoising diffusion probabilistic models (DDPMs) from scratch, with a specific focus on transforming low-resolution microscopy images into their corresponding high-resolution versions. We provide the theoretical background, mathematical derivations, and a detailed Python code implementation using PyTorch, along with techniques to enhance model performance., Comment: 45 pages, 8 figures

In order to relate nanoparticle properties to function, fast and detailed particle characterization, is needed. The ability to characterize nanoparticle samples using optical microscopy techniques has drastically improved over the past few decades; consequently, there are now numerous microscopy methods available for detailed characterization of particles with nanometric size. However, there is currently no ``one size fits all'' solution to the problem of nanoparticle characterization. Instead, since the available techniques have different detection limits and deliver related but different quantitative information, the measurement and analysis approaches need to be selected and adapted for the sample at hand. In this tutorial, we review the optical theory of single particle scattering and how it relates to the differences and similarities in the quantitative particle information obtained from commonly used microscopy techniques, with an emphasis on nanometric (submicron) sized dielectric particles. Particular emphasis is placed on how the optical signal relates to mass, size, structure, and material properties of the detected particles and to its combination with diffusivity-based particle sizing. We also discuss emerging opportunities in the wake of new technology development, with the ambition to guide the choice of measurement strategy based on various challenges related to different types of nanoparticle samples and associated analytical demands.

Critical Casimir forces emerge among particles or surfaces immersed in a near-critical fluid, with the sign of the force determined by surface properties and with its strength tunable by minute temperature changes. Here, we show how such forces can be used to trap a colloidal particle and levitate it above a substrate with a bull's-eye pattern consisting of a ring with surface properties opposite to the rest of the substrate. Using the Derjaguin approximation and mean-field calculations, we find a rich behavior of spherical colloids at such a patterned surface, including sedimentation towards the ring and levitation above the ring (ring levitation) or above the bull's-eye's center (point levitation). Within the Derjaguin approximation, we calculate a levitation diagram for point levitation showing the depth of the trapping potential and the height at which the colloid levitates, both depending on the pattern properties, the colloid size, and the solution temperature. Our calculations reveal that the parameter space associated with point levitation shrinks if the system is driven away from a critical point, while, surprisingly, the trapping force becomes stronger. We discuss the application of critical Casimir levitation for sorting colloids by size and for determining the thermodynamic distance to criticality. Our results show that critical Casimir forces provide rich opportunities for controlling the behavior of colloidal particles at patterned surfaces.

Ellipsoidal particles confined at liquid interfaces exhibit complex self-assembly behaviour due to quadrupolar capillary interactions induced by meniscus deformation. These interactions cause particles to attract each other in either tip-to-tip or side-to-side configurations. However, controlling their interfacial self-assembly is challenging because it is difficult to predict which of these two states will be preferred. In this study, we demonstrate that introducing a soft shell around hard ellipsoidal particles provides a means to control the self-assembly process, allowing us to switch the preferred configuration between these states. We study their interfacial self-assembly and find that pure ellipsoids without a shell consistently form a "chain-like" side-to-side assembly, regardless of aspect ratio. In contrast, core-shell ellipsoids transition from "flower-like" tip-to-tip to "chain-like" side-to-side arrangements as their aspect ratios increase. The critical aspect ratio for transitioning between these structures increases with shell-to-core ratios. Our experimental findings are corroborated by theoretical calculations and Monte Carlo simulations, which map out the phase diagram of thermodynamically preferred self-assembly structures for core-shell ellipsoids as a function of aspect ratio and shell-to-core ratios. This study shows how to program the self-assembly of anisotropic particles by tuning their physicochemical properties, allowing the deterministic realization of distinct structural configurations.

Myxococcus xanthus is a unicellular organism whose cells possess the ability to move and communicate, leading to the emergence of complex collective properties and behaviours. This has made it an ideal model system to study the emergence of collective behaviours in interdisciplinary research efforts lying at the intersection of biology and physics, especially in the growing field of active matter research. Often, challenges arise when setting up reliable and reproducible culturing protocols. This tutorial provides a clear and comprehensive guide on the culture, growth, development, and experimental sample preparation of \textit{M. xanthus}. Additionally, it includes some representative examples of experiments that can be conducted using these samples, namely motility assays, fruiting body formation, predation, and elasticotaxis.

Humanity has long sought inspiration from nature to innovate materials and devices. As science advances, nature-inspired materials are becoming part of our lives. Animate materials, characterized by their activity, adaptability, and autonomy, emulate properties of living systems. While only biological materials fully embody these principles, artificial versions are advancing rapidly, promising transformative impacts across various sectors. This roadmap presents authoritative perspectives on animate materials across different disciplines and scales, highlighting their interdisciplinary nature and potential applications in diverse fields including nanotechnology, robotics and the built environment. It underscores the need for concerted efforts to address shared challenges such as complexity management, scalability, evolvability, interdisciplinary collaboration, and ethical and environmental considerations. The framework defined by classifying materials based on their level of animacy can guide this emerging field encouraging cooperation and responsible development. By unravelling the mysteries of living matter and leveraging its principles, we can design materials and systems that will transform our world in a more sustainable manner.

We introduce optical metasurfaces as components of ultracompact untethered microscopic metaspinners capable of efficient light-induced rotation in a liquid environment. Illuminated by weakly focused light, a metaspinner generates torque via photon recoil through the metasurfaces' ability to bend light towards high angles despite their sub-wavelength thickness, thereby creating orbital angular momentum. We find that a metaspinner is subject to an anomalous transverse optical gradient force that acts in concert with the classical gradient force. Consequently, when two or more metaspinners are trapped together in a laser beam, they collectively orbit the optical axis in the opposite direction to their spinning motion, in stark contrast to rotors coupled through hydrodynamic or mechanical interactions. The metaspinners delineated herein not only serve to illustrate the vast possibilities of utilizing optical metasurfaces for fundamental exploration of optical torques, but they also represent potential building-blocks of artificial active matter systems, light-driven micromachinery, and general-purpose optomechanical devices.

The manipulation of microscopic objects requires precise and controllable forces and torques. Recent advances have led to the use of critical Casimir forces as a powerful tool, which can be finely tuned through the temperature of the environment and the chemical properties of the involved objects. For example, these forces have been used to self-organize ensembles of particles and to counteract stiction caused by Casimir-Liftshitz forces. However, until now, the potential of critical Casimir torques has been largely unexplored. Here, we demonstrate that critical Casimir torques can efficiently control the alignment of microscopic objects on nanopatterned substrates. We show experimentally and corroborate with theoretical calculations and Monte Carlo simulations that circular patterns on a substrate can stabilize the position and orientation of microscopic disks. By making the patterns elliptical, such microdisks can be subject to a torque which flips them upright while simultaneously allowing for more accurate control of the microdisk position. More complex patterns can selectively trap 2D-chiral particles and generate particle motion similar to non-equilibrium Brownian ratchets. These findings provide new opportunities for nanotechnological applications requiring precise positioning and orientation of microscopic objects.

Optical tweezers exploit light--matter interactions to trap particles ranging from single atoms to micrometer-sized eukaryotic cells. For this reason, optical tweezers are a ubiquitous tool in physics, biology, and nanotechnology. Recently, the use of deep learning has started to enhance optical tweezers by improving their design, calibration, and real-time control as well as the tracking and analysis of the trapped objects, often outperforming classical methods thanks to the higher computational speed and versatility of deep learning. Here, we review how deep learning has already remarkably improved optical tweezers, while exploring the exciting, new future possibilities enabled by this dynamic synergy. Furthermore, we offer guidelines on integrating deep learning with optical trapping and optical manipulation in a reliable and trustworthy way., Comment: 19 pages, 7 figures, 1 table

The ability to rapidly detect hydrogen gas upon occurrence of a leak is critical for the safe large-scale implementation of hydrogen (energy) technologies. However, to date, no technically viable sensor solution exists that meets the corresponding response time targets set by stakeholders at technically relevant conditions. Here, we demonstrate how a tailored Long Short-term Transformer Ensemble Model for Accelerated Sensing (LEMAS) accelerates the response of a state-of-the-art optical plasmonic hydrogen sensor by up to a factor of 40 in an oxygen-free inert gas environment, by accurately predicting its response value to a hydrogen concentration change before it is physically reached by the sensor hardware. Furthermore, it eliminates the pressure dependence of the response intrinsic to metal hydride-based sensors, while leveraging their ability to operate in oxygen-starved environments that are proposed to be used for inert gas encapsulation systems of hydrogen installations. Moreover LEMAS provides a measure for the uncertainty of the predictions that is pivotal for safety-critical sensor applications. Our results thus advertise the use of deep learning for the acceleration of sensor response, also beyond the realm of plasmonic hydrogen detection., Comment: 11 pages; 5 figures

The analysis of live-cell single-molecule imaging experiments can reveal valuable information about the heterogeneity of transport processes and interactions between cell components. These characteristics are seen as motion changes in the particle trajectories. Despite the existence of multiple approaches to carry out this type of analysis, no objective assessment of these methods has been performed so far. Here, we have designed a competition to characterize and rank the performance of these methods when analyzing the dynamic behavior of single molecules. To run this competition, we have implemented a software library to simulate realistic data corresponding to widespread diffusion and interaction models, both in the form of trajectories and videos obtained in typical experimental conditions. The competition will constitute the first assessment of these methods, provide insights into the current limits of the field, foster the development of new approaches, and guide researchers to identify optimal tools for analyzing their experiments., Comment: 19 pages, 4 figure, 2 tables. Stage 1 registered report, accepted in principle in Nature Communications (https://springernature.figshare.com/articles/journal_contribution/Quantitative_evaluation_of_methods_to_analyze_motion_changes_in_single-particle_experiments_Registered_Report_Stage_1_Protocol_/24771687)

Unraveling the connections between microscopic structure, emergent physical properties, and slow dynamics has long been a challenge when studying the glass transition. The absence of clear visible structural order in amorphous configurations complicates the identification of the key physical mechanisms underpinning slow dynamics. The difficulty in sampling equilibrated configurations at low temperatures hampers thorough numerical and theoretical investigations. This perspective article explores the potential of machine learning (ML) techniques to face these challenges, building on the algorithms that have revolutionized computer vision and image recognition. We present recent successful ML applications, as well as many open problems for the future, such as transferability and interpretability of ML approaches. We highlight new ideas and directions in which ML could provide breakthroughs to better understand the fundamental mechanisms at play in glass-forming liquids. To foster a collaborative community effort, this article also introduces the ``GlassBench" dataset, providing simulation data and benchmarks for both two-dimensional and three-dimensional glass-formers. We propose critical metrics to compare the performance of emerging ML methodologies, in line with benchmarking practices in image and text recognition. The goal of this roadmap is to provide guidelines for the development of ML techniques in systems displaying slow dynamics, while inspiring new directions to improve our theoretical understanding of glassy liquids.

The implementation of deep learning algorithms has brought new perspectives to plankton ecology. Emerging as an alternative approach to established methods, deep learning offers objective schemes to investigate plankton organisms in diverse environments. We provide an overview of deep-learning-based methods including detection and classification of phyto- and zooplankton images, foraging and swimming behaviour analysis, and finally ecological modelling. Deep learning has the potential to speed up the analysis and reduce the human experimental bias, thus enabling data acquisition at relevant temporal and spatial scales with improved reproducibility. We also discuss shortcomings and show how deep learning architectures have evolved to mitigate imprecise readouts. Finally, we suggest opportunities where deep learning is particularly likely to catalyze plankton research. The examples are accompanied by detailed tutorials and code samples that allow readers to apply the methods described in this review to their own data., Comment: For the associated GitHub repository, see https://github.com/softmatterlab/Deep-learning-in-plankton-ecology

Traditional single-nanoparticle sizing using optical microscopy techniques assesses size via the diffusion constant, which requires suspended particles in a medium of known viscosity. However, these assumptions are typically not fulfilled in complex natural sample environments. Here, we introduce dual-angle interferometric scattering microscopy (DAISY), enabling optical quantification of both size and polarizability of individual nanoparticles without requiring a priori information regarding the surrounding media or super-resolution imaging. DAISY achieves this by combining the information contained in concurrently measured forward and backward scattering images through twilight off-axis holography and interferometric scattering (iSCAT). Going beyond particle size and polarizability, single-particle morphology can be deduced from the fact that hydrodynamic radius relates to the outer particle radius while the scattering-based size estimate depends on the internal mass distribution of the particles. We demonstrate this by optically differentiating biomolecular fractal aggregates from spherical particles in fetal bovine serum at the single particle level.

The Brownian gyrator (BG) is a minimal model of a nano-engine performing a rotational motion, judging solely upon the fact that in non-equilibrium conditions its torque, angular momentum ${\cal L}$ and angular velocity $\cal W$ have non-zero mean values. For a time-discretized model, we calculate the previously unknown probability density functions (PDFs) of ${\cal L}$ and $\cal W$. We find that when the time-step $\delta t \to 0$, both PDFs converge to uniform distributions with diverging variances. For finite $\delta t$, the PDF of ${\cal L}$ has exponential tails and all moments, but its noise-to-signal ratio is generically much bigger than $1$. The PDF of ${\cal W}$ exhibits heavy power-law tails and its mean ${\cal W}$ is the only existing moment. The BG is therefore not an engine in common sense: it does not exhibit regular rotations on each run and its fluctuations are not only a minor nuisance. Our theoretical predictions are confirmed by numerical simulations and experimental data. We discuss some improvements of the model which may result in a more systematic behavior., Comment: 5 pages +SM 11 pages

The formation of groups of interacting individuals improves performance and fitness in many decentralised systems, from micro-organisms to social insects, from robotic swarms to artificial intelligence algorithms. Often, group formation and high-level coordination in these systems emerge from individuals with limited information-processing capabilities implementing low-level rules of communication to signal to each other. Here, we show that, even in a community of clueless individuals incapable of processing information and communicating, a dynamic environment can coordinate group formation by transiently storing memory of the earlier passage of individuals. Our results identify a new mechanism of indirect coordination via shared memory that is primarily promoted and reinforced by dynamic environmental factors, thus overshadowing the need for any form of explicit signalling between individuals. We expect this pathway to group formation to be relevant for understanding and controlling self-organisation and collective decision making in both living and artificial active matter in real-life environments.

Optical tweezers (OT) have become an essential technique in several fields of physics, chemistry, and biology as precise micromanipulation tools and microscopic force transducers. Quantitative measurements require the accurate calibration of the trap stiffness of the optical trap and the diffusion constant of the optically trapped particle. This is typically done by statistical estimators constructed from the position signal of the particle, which is recorded by a digital camera or a quadrant photodiode. The finite integration time and sampling frequency of the detector need to be properly taken into account. Here, we present a general approach based on the joint probability density function of the sampled trajectory that corrects exactly the biases due to the detector's finite integration time and limited sampling frequency, providing theoretical formulas for the most widely employed calibration methods: equipartition, mean squared displacement, autocorrelation, power spectral density, and force reconstruction via maximum-likelihood-estimator analysis (FORMA). Our results, tested with experiments and Monte Carlo simulations, will permit users of OT to confidently estimate the trap stiffness and diffusion constant, extending their use to a broader set of experimental conditions., Comment: 6 figures

Microengines have shown promise for a variety of applications in nanotechnology, microfluidics and nanomedicine, including targeted drug delivery, microscale pumping, and environmental remediation. However, achieving precise control over their dynamics remains a significant challenge. In this study, we introduce a microengine that exploits both optical and thermal effects to achieve a high degree of controllability. We find that in the presence of a strongly focused light beam, a gold-silica Janus particle becomes confined at the stationary point where the optical and thermal forces balance. By using circularly polarized light, we can transfer angular momentum to the particle breaking the symmetry between the two forces and resulting in a tangential force that drives directed orbital motion. We can simultaneously control the velocity and direction of rotation of the particle changing the ellipticity of the incoming light beam, while tuning the radius of the orbit with laser power. Our experimental results are validated using a geometrical optics phenomenological model that considers the optical force, the absorption of optical power, and the resulting heating of the particle. The demonstrated enhanced flexibility in the control of microengines opens up new possibilities for their utilization in a wide range of applications, encompassing microscale transport, sensing, and actuation., Comment: 30 pages, 12 figures

The interaction between metallic and biological nanoparticles (NPs) is widely used in various biotechnology and biomedical applications. However, detailed characterization of this type of interaction is challenging due to a lack of high-throughput techniques that can quantify both size and composition of suspended NP complexes. Here, we introduce a technique called ``twilight nanoparticle tracking analysis'' (tNTA) and demonstrate that it provides a quantitative relationship between the measured optical signal and the composition of suspended dielectric-metal NP complexes. We assess the performance of tNTA by analyzing the selective binding of tannic acid-modified gold nanoparticles (TaAuNPs) to herpes simplex viruses (HSV). Our results show that TaAuNPs bind specifically to HSV without causing substantial changes in the size or refractive index of the virus, suggesting that the binding does not cause virus disruption. Instead, the anti-viral properties of TaAuNPs appear to stem from direct particle binding to the virus.

Through digital imaging, microscopy has evolved from primarily being a means for visual observation of life at the micro- and nano-scale, to a quantitative tool with ever-increasing resolution and throughput. Artificial intelligence, deep neural networks, and machine learning are all niche terms describing computational methods that have gained a pivotal role in microscopy-based research over the past decade. This Roadmap is written collectively by prominent researchers and encompasses selected aspects of how machine learning is applied to microscopy image data, with the aim of gaining scientific knowledge by improved image quality, automated detection, segmentation, classification and tracking of objects, and efficient merging of information from multiple imaging modalities. We aim to give the reader an overview of the key developments and an understanding of possibilities and limitations of machine learning for microscopy. It will be of interest to a wide cross-disciplinary audience in the physical sciences and life sciences.

Adaptivity is a dynamical feature that is omnipresent in nature, socio-economics, and technology. For example, adaptive couplings appear in various real-world systems like the power grid, social, and neural networks, and they form the backbone of closed-loop control strategies and machine learning algorithms. In this article, we provide an interdisciplinary perspective on adaptive systems. We reflect on the notion and terminology of adaptivity in different disciplines and discuss which role adaptivity plays for various fields. We highlight common open challenges, and give perspectives on future research directions, looking to inspire interdisciplinary approaches., Comment: 46 pages, 9 figures

Soft-granular media, such as dense emulsions, foams or tissues, exhibit either fluid- or solid-like properties depending on the applied external stresses. Whereas bulk rheology of such materials has been thoroughly investigated, the internal structural mechanics of finite soft-granular structures with free interfaces is still poorly understood. Here, we report the spontaneous `crystallization' and `melting' inside a model soft granular cluster -- a densely packed aggregate of $N\sim 30-40$ droplets engulfed by a fluid film -- subject to a varying external flow. We develop new machine learning tools to track the internal rearrangements in the quasi-2D cluster as it transits a sequence of constrictions. As the cluster relaxes from a state of strong mechanical deformations, we find differences in the dynamics of the grains within the interior of the cluster and those at its rim, with the latter experiencing larger deformations and less frequent rearrangements, effectively acting as an elastic membrane around a fluid-like core. We conclude that the observed structural-dynamical heterogeneity results from an interplay of the topological constrains, due to the presence of a closed interface, and the internal solid-fluid transitions. We discuss universality of such behavior in various types of finite soft granular structures, including biological tissues., Comment: 16 pages, 9 figures

Light carries energy and momentum. It can therefore alter the motion of objects from atomic to astronomical scales. Being widely available, readily controllable and broadly biocompatible, light is also an ideal tool to propel microscopic particles, drive them out of thermodynamic equilibrium and make them active. Thus, light-driven particles have become a recent focus of research in the field of soft active matter. In this perspective, we discuss recent advances in the control of soft active matter with light, which has mainly been achieved using light intensity. We also highlight some first attempts to utilize light's additional degrees of freedom, such as its wavelength, polarization, and momentum. We then argue that fully exploiting light with all of its properties will play a critical role to increase the level of control over the actuation of active matter as well as the flow of light itself through it. This enabling step will advance the design of soft active matter systems, their functionalities and their transfer towards technological applications.

Preface to the special issue "Characterisation of Physical Processes from Anomalous Diffusion Data" associated with the Anomalous Diffusion Challenge ( https://andi-challenge.org ) and published in Journal of Physics A: Mathematical and Theoretical. The list of articles included in the special issue can be accessed at https://iopscience.iop.org/journal/1751-8121/page/Characterisation-of-Physical-Processes-from-Anomalous-Diffusion-Data ., Comment: Preface to the Special Issue "Characterisation of Physical Processes from Anomalous Diffusion Data", Journal of Physics A: Mathematical and Theoretical https://iopscience.iop.org/journal/1751-8121/page/Characterisation-of-Physical-Processes-from-Anomalous-Diffusion-Data

Specular microscopy assessment of the human corneal endothelium (CE) in Fuchs' dystrophy is challenging due to the presence of dark image regions called guttae. This paper proposes a UNet-based segmentation approach that requires minimal post-processing and achieves reliable CE morphometric assessment and guttae identification across all degrees of Fuchs' dystrophy. We cast the segmentation problem as a regression task of the cell and gutta signed distance maps instead of a pixel-level classification task as typically done with UNets. Compared to the conventional UNet classification approach, the distance-map regression approach converges faster in clinically relevant parameters. It also produces morphometric parameters that agree with the manually-segmented ground-truth data, namely the average cell density difference of -41.9 cells/mm2 (95% confidence interval (CI) [-306.2, 222.5]) and the average difference of mean cell area of 14.8 um2 (95% CI [-41.9, 71.5]). These results suggest a promising alternative for CE assessment.

In the last 20 years, active matter has been a very successful research field, bridging the fundamental physics of nonequilibrium thermodynamics with applications in robotics, biology, and medicine. This field deals with active particles, which, differently from passive Brownian particles, can harness energy to generate complex motions and emerging behaviors. Most active-matter experiments are performed with microscopic particles and require advanced microfabrication and microscopy techniques. Here, we propose some macroscopic experiments with active matter employing commercially available toy robots, i.e., the Hexbugs. We demonstrate how they can be easily modified to perform regular and chiral active Brownian motion. We also show that Hexbugs can interact with passive objects present in their environment and, depending on their shape, set them in motion and rotation. Furthermore, we show that, by introducing obstacles in the environment, we can sort the robots based on their motility and chirality. Finally, we demonstrate the emergence of Casimir-like activity-induced attraction between planar objects in the presence of active particles in the environment., Comment: 11 pages, 5 figures

Optical forces are often calculated by discretizing the trapping light beam into a set of rays and using geometrical optics to compute the exchange of momentum. However, the number of rays sets a trade-off between calculation speed and accuracy. Here, we show that using neural networks permits one to overcome this limitation, obtaining not only faster but also more accurate simulations. We demonstrate this using an optically trapped spherical particle for which we obtain an analytical solution to use as ground truth. Then, we take advantage of the acceleration provided by neural networks to study the dynamics of an ellipsoidal particle in a double trap, which would be computationally impossible otherwise.

Optical tweezers are tools made of light that enable contactless pushing, trapping, and manipulation of objects ranging from atoms to space light sails. Since the pioneering work by Arthur Ashkin in the 1970s, optical tweezers have evolved into sophisticated instruments and have been employed in a broad range of applications in life sciences, physics, and engineering. These include accurate force and torque measurement at the femtonewton level, microrheology of complex fluids, single micro- and nanoparticle spectroscopy, single-cell analysis, and statistical-physics experiments. This roadmap provides insights into current investigations involving optical forces and optical tweezers from their theoretical foundations to designs and setups. It also offers perspectives for applications to a wide range of research fields, from biophysics to space exploration., Comment: 181 pages, 61 figures

Particle tracking is a fundamental task in digital microscopy. Recently, machine-learning approaches have made great strides in overcoming the limitations of more classical approaches. The training of state-of-the-art machine-learning methods almost universally relies on either vast amounts of labeled experimental data or the ability to numerically simulate realistic datasets. However, the data produced by experiments are often challenging to label and cannot be easily reproduced numerically. Here, we propose a novel deep-learning method, named LodeSTAR (Low-shot deep Symmetric Tracking And Regression), that learns to tracks objects with sub-pixel accuracy from a single unlabeled experimental image. This is made possible by exploiting the inherent roto-translational symmetries of the data. We demonstrate that LodeSTAR outperforms traditional methods in terms of accuracy. Furthermore, we analyze challenging experimental data containing densely packed cells or noisy backgrounds. We also exploit additional symmetries to extend the measurable particle properties to the particle's vertical position by propagating the signal in Fourier space and its polarizability by scaling the signal strength. Thanks to the ability to train deep-learning models with a single unlabeled image, LodeSTAR can accelerate the development of high-quality microscopic analysis pipelines for engineering, biology, and medicine., Comment: 19 pages, 4 figures

Casimir forces in quantum electrodynamics emerge between microscopic metallic objects because of the confinement of the vacuum electromagnetic fluctuations occurring even at zero temperature. Their generalization at finite temperature and in material media are referred to as Casimir--Lifshitz forces. These forces are typically attractive, leading to the widespread problem of stiction between the metallic parts of micro- and nanodevices. Recently, repulsive Casimir forces have been experimentally realized but their reliance on specialized materials prevents their dynamic control and thus limits their further applicability. Here, we experimentally demonstrate that repulsive critical Casimir forces, which emerge in a critical binary liquid mixture upon approaching the critical temperature, can be used to actively control microscopic and nanoscopic objects with nanometer precision. We demonstrate this by using critical Casimir forces to prevent the stiction caused by the Casimir--Lifshitz forces. We study a microscopic gold flake above a flat gold-coated substrate immersed in a critical mixture. Far from the critical temperature, stiction occurs because of dominant Casimir--Lifshitz forces. Upon approaching the critical temperature, however, we observe the emergence of repulsive critical Casimir forces that are sufficiently strong to counteract stiction. This experimental demonstration can accelerate the development of micro- and nanodevices by preventing stiction as well as providing active control and precise tunability of the forces acting between their constituent parts., Comment: 27 pages, 5 figures

The marine microbial food web plays a central role in the global carbon cycle. Our mechanistic understanding of the ocean, however, is biased towards its larger constituents, while rates and biomass fluxes in the microbial food web are mainly inferred from indirect measurements and ensemble averages. Yet, resolution at the level of the individual microplankton is required to advance our understanding of the oceanic food web. Here, we demonstrate that, by combining holographic microscopy with deep learning, we can follow microplanktons throughout their lifespan, continuously measuring their three dimensional position and dry mass. The deep learning algorithms circumvent the computationally intensive processing of holographic data and allow rapid measurements over extended time periods. This permits us to reliably estimate growth rates, both in terms of dry mass increase and cell divisions, as well as to measure trophic interactions between species such as predation events. The individual resolution provides information about selectivity, individual feeding rates and handling times for individual microplanktons. This method is particularly useful to explore the flux of carbon through micro-zooplankton, the most important and least known group of primary consumers in the global oceans. We exemplify this by detailed descriptions of micro-zooplankton feeding events, cell divisions, and long term monitoring of single cells from division to division., Comment: 20 pages, 4 figure, 5 supplementary figure

The characterization of dynamical processes in living systems provides important clues for their mechanistic interpretation and link to biological functions. Thanks to recent advances in microscopy techniques, it is now possible to routinely record the motion of cells, organelles, and individual molecules at multiple spatiotemporal scales in physiological conditions. However, the automated analysis of dynamics occurring in crowded and complex environments still lags behind the acquisition of microscopic image sequences. Here, we present a framework based on geometric deep learning that achieves the accurate estimation of dynamical properties in various biologically-relevant scenarios. This deep-learning approach relies on a graph neural network enhanced by attention-based components. By processing object features with geometric priors, the network is capable of performing multiple tasks, from linking coordinates into trajectories to inferring local and global dynamic properties. We demonstrate the flexibility and reliability of this approach by applying it to real and simulated data corresponding to a broad range of biological experiments., Comment: 17 pages, 5 figure, 2 supplementary figures

Active matter comprises self-driven units, such as bacteria and synthetic microswimmers, that can spontaneously form complex patterns and assemble into functional microdevices. These processes are possible thanks to the out-of-equilibrium nature of active-matter systems, fueled by a one-way free-energy flow from the environment into the system. Here, we take the next step in the evolution of active matter by realizing a two-way coupling between active particles and their environment, where active particles act back on the environment giving rise to the formation of superstructures. In experiments and simulations we observe that, under light-illumination, colloidal particles and their near-critical environment create mutually-coupled co-evolving structures. These structures unify in the form of active superstructures featuring a droplet shape and a colloidal engine inducing self-propulsion. We call them active droploids -- a portmanteau of droplet and colloids. Our results provide a pathway to create active superstructures through environmental feedback.

Neural network training and validation rely on the availability of large high-quality datasets. However, in many cases only incomplete datasets are available, particularly in health care applications, where each patient typically undergoes different clinical procedures or can drop out of a study. Since the data to train the neural networks need to be complete, most studies discard the incomplete datapoints, which reduces the size of the training data, or impute the missing features, which can lead to artefacts. Alas, both approaches are inadequate when a large portion of the data is missing. Here, we introduce GapNet, an alternative deep-learning training approach that can use highly incomplete datasets. First, the dataset is split into subsets of samples containing all values for a certain cluster of features. Then, these subsets are used to train individual neural networks. Finally, this ensemble of neural networks is combined into a single neural network whose training is fine-tuned using all complete datapoints. Using two highly incomplete real-world medical datasets, we show that GapNet improves the identification of patients with underlying Alzheimer's disease pathology and of patients at risk of hospitalization due to Covid-19. By distilling the information available in incomplete datasets without having to reduce their size or to impute missing values, GapNet will permit to extract valuable information from a wide range of datasets, benefiting diverse fields from medicine to engineering., Comment: 11 pages, 3 figures, 1 table

Deviations from Brownian motion leading to anomalous diffusion are ubiquitously found in transport dynamics, playing a crucial role in phenomena from quantum physics to life sciences. The detection and characterization of anomalous diffusion from the measurement of an individual trajectory are challenging tasks, which traditionally rely on calculating the mean squared displacement of the trajectory. However, this approach breaks down for cases of important practical interest, e.g., short or noisy trajectories, ensembles of heterogeneous trajectories, or non-ergodic processes. Recently, several new approaches have been proposed, mostly building on the ongoing machine-learning revolution. Aiming to perform an objective comparison of methods, we gathered the community and organized an open competition, the Anomalous Diffusion challenge (AnDi). Participating teams independently applied their own algorithms to a commonly-defined dataset including diverse conditions. Although no single method performed best across all scenarios, the results revealed clear differences between the various approaches, providing practical advice for users and a benchmark for developers., Comment: 63 pages, 5 main figures, 1 table, 28 supplementary figures. Website: http://www.andi-challenge.org

Countless systems in biology, physics, and finance undergo diffusive dynamics. Many of these systems, including biomolecules inside cells, active matter systems and foraging animals, exhibit anomalous dynamics where the growth of the mean squared displacement with time follows a power law with an exponent that deviates from $1$. When studying time series recording the evolution of these systems, it is crucial to precisely measure the anomalous exponent and confidently identify the mechanisms responsible for anomalous diffusion. These tasks can be overwhelmingly difficult when only few short trajectories are available, a situation that is common in the study of non-equilibrium and living systems. Here, we present a data-driven method to analyze single anomalous diffusion trajectories employing recurrent neural networks, which we name RANDI. We show that our method can successfully infer the anomalous exponent, identify the type of anomalous diffusion process, and segment the trajectories of systems switching between different behaviors. We benchmark our performance against the state-of-the art techniques for the study of single short trajectories that participated in the Anomalous Diffusion (AnDi) Challenge. Our method proved to be the most versatile method, being the only one to consistently rank in the top 3 for all tasks proposed in the AnDi Challenge.

Quantitative analysis of cell structures is essential for biomedical and pharmaceutical research. The standard imaging approach relies on fluorescence microscopy, where cell structures of interest are labeled by chemical staining techniques. However, these techniques are often invasive and sometimes even toxic to the cells, in addition to being time-consuming, labor-intensive, and expensive. Here, we introduce an alternative deep-learning-powered approach based on the analysis of brightfield images by a conditional generative adversarial neural network (cGAN). We show that this approach can extract information from the brightfield images to generate virtually-stained images, which can be used in subsequent downstream quantitative analyses of cell structures. Specifically, we train a cGAN to virtually stain lipid droplets, cytoplasm, and nuclei using brightfield images of human stem-cell-derived fat cells (adipocytes), which are of particular interest for nanomedicine and vaccine development. Subsequently, we use these virtually-stained images to extract quantitative measures about these cell structures. Generating virtually-stained fluorescence images is less invasive, less expensive, and more reproducible than standard chemical staining; furthermore, it frees up the fluorescence microscopy channels for other analytical probes, thus increasing the amount of information that can be extracted from each cell., Comment: 12 pages, 4 figures

Nanostructured dielectric metasurfaces offer unprecedented opportunities to manipulate light by imprinting an arbitrary phase-gradient on an impinging wavefront. This has resulted in the realization of a range of flat analogs to classical optical components like lenses, waveplates and axicons. However, the change in linear and angular optical momentum associated with phase manipulation also results in previously unexploited forces acting on the metasurface itself. Here, we show that these optomechanical effects can be utilized to construct optical metavehicles - microscopic particles that can travel long distances under low-power plane-wave illumination while being steered through the polarization of the incident light. We demonstrate movement in complex patterns, self-correcting motion, and an application as transport vehicles for microscopic cargo, including unicellular organisms. The abundance of possible optical metasurfaces attests to the prospect of developing a wide variety of metavehicles with specialized functional behavior., Comment: Main Text: 16 pages, 4 figures. Supporting Information: 6 pages, 9 figures

Containment of epidemic outbreaks entails great societal and economic costs. Cost-effective containment strategies rely on efficiently identifying infected individuals, making the best possible use of the available testing resources. Therefore, quickly identifying the optimal testing strategy is of critical importance. Here, we demonstrate that machine learning can be used to identify which individuals are most beneficial to test, automatically and dynamically adapting the testing strategy to the characteristics of the disease outbreak. Specifically, we simulate an outbreak using the archetypal susceptible-infectious-recovered (SIR) model and we use data about the first confirmed cases to train a neural network that learns to make predictions about the rest of the population. Using these prediction, we manage to contain the outbreak more effectively and more quickly than with standard approaches. Furthermore, we demonstrate how this method can be used also when there is a possibility of reinfection (SIRS model) to efficiently eradicate an endemic disease., Comment: 11 pages, 4 figures

Video microscopy has a long history of providing insights and breakthroughs for a broad range of disciplines, from physics to biology. Image analysis to extract quantitative information from video microscopy data has traditionally relied on algorithmic approaches, which are often difficult to implement, time consuming, and computationally expensive. Recently, alternative data-driven approaches using deep learning have greatly improved quantitative digital microscopy, potentially offering automatized, accurate, and fast image analysis. However, the combination of deep learning and video microscopy remains underutilized primarily due to the steep learning curve involved in developing custom deep-learning solutions. To overcome this issue, we introduce a software, DeepTrack 2.0, to design, train and validate deep-learning solutions for digital microscopy. We use it to exemplify how deep learning can be employed for a broad range of applications, from particle localization, tracking and characterization to cell counting and classification. Thanks to its user-friendly graphical interface, DeepTrack 2.0 can be easily customized for user-specific applications, and, thanks to its open-source object-oriented programming, it can be easily expanded to add features and functionalities, potentially introducing deep-learning-enhanced video microscopy to a far wider audience., Comment: 25 pages; 13 figures

Active particles break out of thermodynamic equilibrium thanks to their directed motion, which leads to complex and interesting behaviors in the presence of confining potentials. When dealing with active nanoparticles, however, the overwhelming presence of rotational diffusion hinders directed motion, leading to an increase of their effective temperature, but otherwise masking the effects of self-propulsion. Here, we demonstrate an experimental system where an active nanoparticle immersed in a critical solution and held in an optical harmonic potential features far-from-equilibrium behavior beyond an increase of its effective temperature. When increasing the laser power, we observe a cross-over from a Boltzmann distribution to a non-equilibrium state, where the particle performs fast orbital rotations about the beam axis. These findings are rationalized by solving the Fokker-Planck equation for the particle's position and orientation in terms of a moment expansion. The proposed self-propulsion mechanism results from the particle's non-sphericity and the lower critical point of the solute., Comment: 6 figures

Critical Casimir forces emerge between objects, such as colloidal particles, whenever their surfaces spatially confine the fluctuations of the order parameter of a critical liquid used as a solvent. These forces act at short but microscopically large distances between these objects, reaching often hundreds of nanometers. Keeping colloids at such distances is a major experimental challenge, which can be addressed by the means of optical tweezers. Here, we review how optical tweezers have been successfully used to quantitatively study critical Casimir forces acting on particles in suspensions. As we will see, the use of optical tweezers to experimentally study critical Casimir forces can play a crucial role in developing nano-technologies, representing an innovative way to realize self-assembled devices at the nano- and microscale., Comment: 18 pages, 11 figures