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Patients with colorectal liver metastasis (CLM) present with heterogenous clinical outcomes and improved classification is needed to ameliorate the therapeutic output. Macrophages (Mϕ) hold promise as prognostic classifiers and therapeutic targets. Here, stemming from a single-cell analysis of mononuclear phagocytes infiltrating human CLM, we identified two Mϕ markers associated with distinct populations with opposite clinical relevance. The invasive margin of CLM was enriched in pro-inflammatory monocyte-derived Mϕ (MoMϕ) expressing the monocytic marker SERPINB2, and a more differentiated population, tumor-associated Mϕ (TAM), expressing glycoprotein nonmetastatic melanoma protein B (GPNMB). SERPINB2+ MoMϕ had an early inflammatory profile, whereas GPNMB+ TAMs were enriched in pathways of matrix degradation, angiogenesis, and lipid metabolism and were found closer to the tumor margin, as confirmed by spatial transcriptomics on CLM specimens. In a cohort of patients, a high infiltration of SERPINB2+ cells independently associated with longer disease-free survival (DFS; P = 0.033), whereas a high density of GPNMB+ cells correlated with shorter DFS (P = 0.012) and overall survival (P = 0.002). Cell–cell interaction analysis defined opposing roles for MoMϕ and TAMs, suggesting that SERPINB2+ and GPNMB+ cells are discrete populations of Mϕ and may be exploited for further translation to an immune-based stratification tool. This study provides evidence of how multi-omics approaches can identify nonredundant, clinically relevant markers for further translation to immune-based patient stratification tools and therapeutic targets. GPNMB has been shown to set Mϕ in an immunosuppressive mode. Our high dimensional analyses provide further evidence that GPNMB is a negative prognostic indicator and a potential player in the protumor function of Mϕ populations.

Image sequences of brownian colloidal particles and flowing blood cells behind parafilm layers. Code for the analysis and inversion of the mean square displacement.

Supplementary Figures 1-4, Tables S1 and S5

Supplementary Table S4

Patients with colorectal liver metastasis (CLM) present with heterogenous clinical outcomes and improved classification is needed to ameliorate the therapeutic output. Macrophages (Mϕ) hold promise as prognostic classifiers and therapeutic targets. Here, stemming from a single-cell analysis of mononuclear phagocytes infiltrating human CLM, we identified two Mϕ markers associated with distinct populations with opposite clinical relevance. The invasive margin of CLM was enriched in pro-inflammatory monocyte-derived Mϕ (MoMϕ) expressing the monocytic marker SERPINB2, and a more differentiated population, tumor-associated Mϕ (TAM), expressing glycoprotein nonmetastatic melanoma protein B (GPNMB). SERPINB2+ MoMϕ had an early inflammatory profile, whereas GPNMB+ TAMs were enriched in pathways of matrix degradation, angiogenesis, and lipid metabolism and were found closer to the tumor margin, as confirmed by spatial transcriptomics on CLM specimens. In a cohort of patients, a high infiltration of SERPINB2+ cells independently associated with longer disease-free survival (DFS; P = 0.033), whereas a high density of GPNMB+ cells correlated with shorter DFS (P = 0.012) and overall survival (P = 0.002). Cell–cell interaction analysis defined opposing roles for MoMϕ and TAMs, suggesting that SERPINB2+ and GPNMB+ cells are discrete populations of Mϕ and may be exploited for further translation to an immune-based stratification tool. This study provides evidence of how multi-omics approaches can identify nonredundant, clinically relevant markers for further translation to immune-based patient stratification tools and therapeutic targets. GPNMB has been shown to set Mϕ in an immunosuppressive mode. Our high dimensional analyses provide further evidence that GPNMB is a negative prognostic indicator and a potential player in the protumor function of Mϕ populations.

Supplementary figure legends

Abstract The connection between the properties of a cell tissue and those of the single constituent cells remains to be elucidated. At the purely mechanical level, the degree of rigidity of different cellular components, such as the nucleus and the cytoplasm, modulates the interplay between the cell inner processes and the external environment, while simultaneously mediating the mechanical interactions between neighboring cells. Being able to quantify the correlation between single-cell and tissue properties would improve our mechanobiological understanding of cell tissues. Here we develop a methodology to quantitatively extract a set of structural and motility parameters from the analysis of time-lapse movies of nuclei belonging to jammed and flocking cell monolayers. We then study in detail the correlation between the dynamical state of the tissue and the deformation of the nuclei. We observe that the nuclear deformation rate linearly correlates with the local divergence of the velocity field, which leads to a non-invasive estimate of the elastic modulus of the nucleus relative to the one of the cytoplasm. We also find that nuclei belonging to flocking monolayers, subjected to larger mechanical perturbations, are about two time stiffer than nuclei belonging to dynamically arrested monolayers, in agreement with atomic force microscopy results. Our results demonstrate a non-invasive route to the determination of nuclear relative stiffness for cells in a monolayer. Graphic abstract

On approaching the glass transition, a dense colloid exhibits a dramatic slowdown with minute structural changes. Most microscopy experiments directly follow the motion of individual particles in real space, whereas scattering experiments typically probe the collective dynamics in reciprocal space at variable wavevector q. Multiscale studies of glassy dynamics are experimentally demanding and, thus, seldom performed. By using two-dimensional hard-sphere colloids at various area fractions ϕ, we show here that Differential Dynamic Microscopy (DDM) can be effectively used to measure the collective dynamics of a glassy colloid in a range of q within a single experiment. As ϕ is increased, the single decay of the intermediate scattering functions is progressively replaced by a more complex relaxation that we fit to a sum of two stretched-exponential decays. The slowest process, corresponding to the long-time particle escapes from caging, has a characteristic time τ s = 1/( D L q2) with diffusion coefficient [Formula: see text], and ϕ c ≃ 0.81. The fast process exhibits, instead, a non-Brownian scaling of the characteristic time τ f( q) and a relative amplitude a( q) that monotonically increases with q. Despite the non-Brownian nature of τ f( q), we succeed in estimating the short-time diffusion coefficient D cage, whose ϕ-dependence is practically negligible compared to the one of D L. Finally, we extend DDM to measure the q-dependent dynamical susceptibility χ4( q, t), a powerful yet hard-to-access multiscale indicator of dynamical heterogeneities. Our results show that DDM is a convenient tool to study the dynamics of colloidal glasses over a broad range of time and length scales.

The process in which locally confined epithelial malignancies progressively evolve into invasive cancers is often promoted by unjamming, a phase transition from a solid-like to a liquid-like state that occurs in various tissues. Whether this tissue-level mechanical transition impact phenotypes during carcinoma progression remains unclear. We show, here that the large fluctuations in cell density that accompany unjamming result in repeated mechanical deformations of cells and nuclei. Cells react to these protracted mechanical stresses by mounting a mechano-protective response that includes enlarged nuclear size and rigidity, altered heterochromatin distribution, and the remodeling of the perinuclear actin architecture into actin rings. The chronic strains and stresses associated with unjamming together with reduction of Lamin B1 levels eventually result in DNA damage and nuclear envelope ruptures, with the release of cytosolic DNA that activates a cGAS/STING-dependent cytosolic DNA response gene program. This mechanically-driven transcriptional rewiring ultimately results in a change in cell state, with the emergence of malignant traits, including epithelial-to-mesenchymal plasticity phenotypes and chemo-resistance in invasive breast carcinoma. One-Sentence Summary: A solid-to-fluid phase transition promotes a pro-inflammatory transcriptional response in invasive breast carcinoma

Abstract Soft and biological materials are often composed of elementary constituents exhibiting an incessant roto-translational motion at the microscopic scale. Tracking this motion with a bright-field microscope becomes increasingly challenging when the particle size becomes smaller than the microscope resolution, a case which is frequently encountered. Here we demonstrate squared-gradient differential dynamic microscopy (SG-DDM) as a tool to successfully use bright-field microscopy to extract the roto-translational dynamics of small anisotropic colloidal particles, whose rotational motion cannot be tracked accurately in direct space. We provide analytical justification and experimental demonstration of the method by successful application to an aqueous suspension of peanut-shaped particles. Graphic abstract

Specialised ribonucleoprotein (RNP) granules are a hallmark of polarized cells, like neurons and germ cells. Among their main functions is the spatial and temporal modulation of the activity of specific mRNA transcripts that allow specification of primary embryonic axes. While RNPs composition and role are well established, their regulation is poorly defined. Here, we demonstrate that Hecw, a newly identified Drosophila ubiquitin ligase, is a key modulator of RNPs in oogenesis and neurons. Hecw depletion leads to the formation of enlarged granules that transition from a liquid to a gel-like state. Loss of Hecw activity results in defective oogenesis, premature aging and climbing defects associated with neuronal loss. At the molecular level, reduced ubiquitination of the Fmrp impairs its translational repressor activity, resulting in altered Orb expression in nurse cells and Profilin in neurons., Ribonucleoprotein (RNP) granules are responsible for mRNA transport and local translation required for neuronal and oocyte maturation. Here the authors show that loss of the Drosophila Ub ligase Hecw enlarges RNP granules, leads to a liquid to gel-like transition, and results in defective oogenesis and neuronal loss.

Oscillatory shear tests are widely used in rheology to characterize the linear and non-linear mechanical response of complex fluids, including the yielding transition. There is an increasing urge to acquire detailed knowledge of the deformation field that is effectively present across the sample during these tests; at the same time, there is mounting evidence that the macroscopic rheological response depends on the elusive microscopic behavior of the material constituents. Here we employ a strain-controlled shear-cell with transparent walls to visualize and quantify the dynamics of tracers embedded in various cyclically sheared complex fluids, ranging from almost-ideal elastic to yield stress fluids. For each sample, we use image correlation processing to measure the macroscopic deformation field, and echo-differential dynamic microscopy to probe the microscopic irreversible sample dynamics in reciprocal space; finally, we devise a simple scheme to spatially map the rearrangements in the sheared sample, once again without tracking the tracers. For the yield stress sample, we obtain a wave-vector dependent characterization of shear-induced diffusion across the yielding transition, which is accompanied by a three-order-of-magnitude speed-up of the dynamics and by a transition from localized, intermittent rearrangements to a more spatially homogeneous and temporally uniform activity. Our tracking free approach is intrinsically multi-scale, can successfully discriminate between different types of dynamics, and can be automated to minimize user intervention. Applications are many, as it can be translated to other imaging modes, including fluorescence, and can be used with sub-resolution tracers and even without tracers, for samples that provide intrinsic optical contrast.

This review summarizes recent progress in investigating polymer systems by using Differential dynamic microscopy (DDM), a rapidly emerging approach that transforms a commercial microscope by combining real-space information with the powerful capabilities of conventional light scattering analysis. DDM analysis of a single microscope movie gives access to the sample dynamics in a wide range of scattering wave-vectors, enabling contemporary polymer science experiments that would be difficult or impossible with standard light scattering techniques. Examples of application include the characterization of polymer solutions and networks, of polymer based colloidal systems, of biopolymers, and of cellular motility in polymeric fluids. Further applications of DDM to a variety of polymer systems are suggested to be just behind the corner and it is thus likely that DDM will become a tool of choice of the modern experimental polymer scientists.

The accurate quantification of cellular motility and of the structural changes occurring in multicellular aggregates is critical in describing and understanding key biological processes, such as wound repair, embryogenesis and cancer invasion. Current methods based on cell tracking or velocimetry either suffer from limited spatial resolution or are challenging and time-consuming, especially for three-dimensional (3D) cell assemblies. Here we propose a conceptually simple, robust and tracking-free approach for the quantification of the dynamical activity of cells via a two-step procedure. We first characterise the global features of the collective cell migration by registering the temporal stack of the acquired images. As a second step, a map of the local cell motility is obtained by performing a mean squared amplitude analysis of the intensity fluctuations occurring when two registered image frames acquired at different times are subtracted. We successfully apply our approach to cell monolayers undergoing a jamming transition, as well as to monolayers and 3D aggregates that exhibit a cooperative unjamming-via-flocking transition. Our approach is capable of disentangling very efficiently and of assessing accurately the global and local contributions to cell motility.

The GRADFLEX experiment of ESA has shown that under microgravity conditions a stationary thermodiffusion process is accompanied by giant non-equilibrium fluctuations with size as large as the size of the sample. In the presence of small concentration gradients, the features of the non-equilibrium fluctuations can be described by means of linearized hydrodynamics. However, the linear models are not suitable to describe most cases of applicative interest, such as fluctuations induced by large gradients and under non-stationary conditions. Moreover, presently the investigation of non-equilibrium fluctuations has mainly involved single component fluids and binary mixtures, but recently transport processes in ternary mixtures have attracted increasing interest due to the experiments performed on the International Space Station in the framework of the DCMIX project of ESA. The Giant Fluctuations (NEUF-DIX) project of ESA will investigate non-equilibrium fluctuations during diffusive processes occurring in complex multi-component mixtures, where one of the components is a macromolecule, such as a polymer, a colloid or a protein. Important objectives will be the exploration of the features of the fluctuations under non-ideal conditions, such as large gradients, transient processes, and concentrated samples, and the understanding of how the fluctuations affect the interactions between macromolecules. The project involves the development of a dedicated facility, consisting of an array of shadowgraph optical instruments working in parallel, each one equipped with a thermal gradient cell. Here we outline the design concept of the facility and the results of performance tests performed on a breadboard to evaluate the suitability of the designed instrument to carry out scientific measurements of non-equilibrium concentration fluctuations in space.

We use differential dynamic microscopy and particle tracking to determine the dynamical characteristics of a coarsening foam in reciprocal and direct space. At all wavevectors $q$ investigated, the intermediate scattering function exhibits a compressed exponential decay. However, the access to unprecedentedly small $q$s highlights the existence of two distinct regimes for the $q$-dependence of the foam relaxation rate $\Gamma (q)$. At any given foam age, $\Gamma (q)\sim q$ at high $q$, consistent with directionally-persistent and intermittent bubble displacements. At low $q$, we find $\Gamma (q) \sim q^{1.6}$. We show that such change in $q$-dependence of $\Gamma (q)$ relates to a bubble displacement distribution exhibiting a cut-off length of the order of the bubble diameter. Investigations of the $q$-dependence of $\Gamma (q)$ at different foam ages reveal that foam dynamics is not only governed by the bubble length scale, but also by the strain rate imposed by the bubble growth; normalizing $\Gamma (q)$ by this strain rate and multiplying $q$ with the age-dependent bubble radius leads to a collapse of all data sets onto a unique master-curve., Comment: J. Phys.: Condens. Matter (2020)

Specialised ribonucleoprotein (RNP) granules are a hallmark of germ cells. Among their main function is the spatial and temporal modulation of the activity of specific mRNA transcripts that allow specification of primary embryonic axes. While RNPs composition and role are well established, their regulation is poorly defined. Here, we demonstrate that Hecw, a newly identifiedDrosophilaubiquitin ligase, is a key modulator of RNPs in oogenesis. Loss of Hecw activity results in the formation of enlarged granules that transition from a liquid to a gel-like state. At the molecular level, Hecw depletion leads to reduced ubiquitination and activity of the translational repressor Fmrp, resulting in premature Orb expression/recruitment in nurse cells. In addition to defective oogenesis, flies lacking Hecw show neurodegenerative traits with premature aging and climbing defects due to neuronal loss that are linked to RNPs condensation. Our findings reveal an unprecedented function of ubiquitin in modulating RNP fluidity and activity.

Transport in cells occurs via a delicate interplay of passive and active processes, including diffusion, directed transport and advection. Despite progress in super-resolution microscopy, discriminating and quantifying these processes is a challenge, requiring tracking of rapidly moving, sub-diffraction objects in a crowded, noisy environment. Here we use differential dynamic microscopy with different contrast mechanisms to provide a thorough characterization of the dynamics in the Drosophila oocyte. We study the movement of vesicles and the elusive motion of a cytoplasmic F-actin mesh, a known regulator of cytoplasmic flows. We find that cytoplasmic motility constitutes a combination of directed motion and random diffusion. While advection is mainly attributed to microtubules, we find that active diffusion is driven by the actin cytoskeleton, although it is also enhanced by the flow. We also find that an important dynamic link exists between vesicles and cytoplasmic F-actin motion, as recently suggested in mouse oocytes., Intracellular transport is facilitated by a combination of processes including directed transport, advection and diffusion. Here the authors microscopically characterise the dynamics of the Drosophila oocyte and find distinct contributions of cytoskeletal components to advection and active diffusion.

This paper describes the European Space Agency (ESA) experiments devoted to study thermodiffusion of fluid mixtures in microgravity environment, where sedimentation and convection do not affect the mass flow induced by the Soret effect. First, the experiments performed on binary mixtures in the IVIDIL and GRADFLEX experiments are described. Then, further experiments on ternary mixtures and complex fluids performed in DCMIX and planned to be performed in the context of the NEUF-DIX project are presented. Finally, multi-component mixtures studied in the SCCO project are detailed. Graphical abstract: [Figure not available: see fulltext.]., 0, SCOPUS: re.j, info:eu-repo/semantics/published

Damage-induced long non-coding RNAs (dilncRNA) synthesized at DNA double-strand breaks (DSBs) by RNA polymerase II (RNAPII) are necessary for DNA damage response (DDR) foci formation. We demonstrate that induction of DSBs results in the assembly of functional promoters that include a complete RNAPII pre-initiation complex (PIC), MED1 and CDK9. Absence or inactivation of these factors causes DDR foci reduction both in vivo and in an in vitro system that reconstitutes DDR events on nucleosomes. We also show that dilncRNAs drive molecular crowding of DDR proteins such as 53BP1 into foci that exhibit liquid-liquid phase separation (LLPS) condensate properties. We propose that the assembly of DSB-induced transcriptional promoters drives RNA synthesis which stimulates phase separation of DDR factors in the shape of foci.

Under homeostatic conditions, mature epithelia are locked in a kinetically-silent, jammed state. During wound repair or branching morphogenesis epithelia must unjam and acquire liquid-like properties. These events might be recapitulated in the transition from in situ to invasive cancer stages. How cells control this transition and how biologically relevant it is, however, remains unclear. Recently, we showed that altering RAB5A levels, a master regulator of endosomal trafficking, is sufficient to re-awaken motility in jammed epithelia, through ill-defined, endocytic-sensitive biochemical pathways. Here, we show that RAB5A promotes non-clathrin-dependent internalization of epidermal growth factor receptor that leads to the hyper-activation of endosomally-confined ERK1/2 and phosphorylation of the branched actin nucleator, WAVE2. This endo-ERK1/2 route, in turn, is critical to promote a flocking transition in epithelial monolayers and in differentiated normal mammary 3D cysts, where it further overcome proliferation arrest and to initiate bud morphogenesis. In 3D spheroid models of breast ductal carcinoma in situ (DCIS), instead, endo-ERK1/2-mediated unjamming causes the emergence of highly coordinated angular rotational motion, matrix remodelling and collective 3D invasion. Similarly, it promotes a flocking mode of collective dissemination ex vivo in slices of orthotopically-implanted DCIS. Thus, EGF-dependent activation of endosomal ERK1/2 is found to be the molecular determinant of unjamming via flocking transition: a powerful machinery that overcomes the kinetic and proliferation arrest of terminally differentiated epithelial cells, and promotes collective invasive programs of jammed carcinomas.

During wound repair, branching morphogenesis and carcinoma dissemination, cellular rearrangements are fostered by a solid-to-liquid transition, known as unjamming. The biomolecular machinery behind unjamming and its pathophysiological relevance remain, however, unclear. Here, we study unjamming in a variety of normal and tumorigenic epithelial two-dimensional (2D) and 3D collectives. Biologically, the increased level of the small GTPase RAB5A sparks unjamming by promoting non-clathrin-dependent internalization of epidermal growth factor receptor that leads to hyperactivation of the kinase ERK1/2 and phosphorylation of the actin nucleator WAVE2. This cascade triggers collective motility effects with striking biophysical consequences. Specifically, unjamming in tumour spheroids is accompanied by persistent and coordinated rotations that progressively remodel the extracellular matrix, while simultaneously fluidizing cells at the periphery. This concurrent action results in collective invasion, supporting the concept that the endo-ERK1/2 pathway is a physicochemical switch to initiate collective invasion and dissemination of otherwise jammed carcinoma. A RAB5A-mediated, epidermal growth factor-dependent activation of endosomal ERK1/2 is identified as a key molecular route for a solid-to-liquid-like phase transition, sufficient to overcome kinetic and proliferation arrest in normal mammary epithelial assemblies and to promote collective invasion in breast carcinoma.

Collective cell migration in dense tissues underlies important biological processes, such as embryonic development, wound healing and cancer invasion. While many aspects of single cell movements are now well established, the mechanisms leading to displacements of cohesive cell groups are still poorly understood. To elucidate the emergence of collective migration in mechanosensitive cells, we examine a self-propelled Voronoi (SPV) model of confluent tissues with an orientational feedback that aligns a cell's polarization with its local migration velocity. While shape and motility are known to regulate a density-independent liquid–solid transition in tissues, we find that aligning interactions facilitate collective motion and promote solidification, with transitions that can be predicted by extending statistical physics tools such as effective temperature to this far-from-equilibrium system. In addition to accounting for recent experimental observations obtained with epithelial monolayers, our model predicts structural and dynamical signatures of flocking, which may serve as gateway to a more quantitative characterization of collective motility.

Differential Dynamic Microscopy (DDM) analyzes traditional real-space microscope images to extract information on sample dynamics in a way akin to light scattering, by decomposing each image in a sequence into Fourier modes, and evaluating their time correlation properties. DDM has been applied in a number of soft-matter and colloidal systems. However, objects observed to move out of the microscope's captured field of view, intersecting the edges of the acquired images, can introduce spurious but significant errors in the subsequent analysis. Here we show that application of a spatial windowing filter to images in a sequence before they enter the standard DDM analysis can reduce these artifacts substantially. Moreover, windowing can increase significantly the accessible range of wave vectors probed by DDM, and may further yield unexpected information, such as the size polydispersity of a colloidal suspension.

Micro- and nanoscale objects with anisotropic shape are key components of a variety of biological systems and inert complex materials, and represent fundamental building blocks of novel self-assembly strategies. The time scale of their thermal motion is set by their translational and rotational diffusion coefficients, whose measurement may become difficult for relatively large particles with small optical contrast. Here we show that dark field differential dynamic microscopy is the ideal tool for probing the roto-translational Brownian motion of anisotropic shaped particles. We demonstrate our approach by successful application to aqueous dispersions of non-motile bacteria and of colloidal aggregates of spherical particles.

Particle tracking microrheology (PT-$\mu$r) exploits the thermal motion of embedded particles to probe the local mechanical properties of soft materials. Despite its appealing conceptual simplicity, PT-$\mu$r requires calibration procedures and operating assumptions that constitute a practical barrier to a wider adoption. Here we demonstrate Differential Dynamic Microscopy microrheology (DDM-$\mu$r), a tracking-free approach based on the multi-scale, temporal correlation study of the image intensity fluctuations that are observed in microscopy experiments as a consequence of the motion of the tracers. We show that the mechanical moduli of an arbitrary sample are determined correctly in a wide frequency range, provided that the standard DDM analysis is reinforced with a novel, iterative, self-consistent procedure that fully exploits the multi-scale information made available by DDM. Our approach to DDM-$\mu$r does not require any prior calibration, is in agreement with both traditional rheology and Diffusing Wave Spectroscopy microrheology, and works in conditions where PT-$\mu$r fails, providing thus an operationally simple, calibration-free probe of soft materials., Comment: 10 pages, 5 figures

Transport in cells occurs via a delicate interplay of passive and active processes, including diffusion, directed transport and advection. Despite progresses in super-resolution microscopy, discriminating and quantifying these processes is a challenge, requiring tracking of rapidly moving, sub-diffraction objects in a crowded, noisy environment. Here we use Differential Dynamic Microscopy with different contrast mechanisms to provide a thorough characterization of the dynamics in theDrosophilaoocyte. We study the movement of vesicles and the elusive motion of a cytoplasmic F-actin mesh, a known regulator of cytoplasmic flows. We find that cytoplasmic motility constitutes a combination of directed motion and random diffusion. While advection is mainly attributed to microtubules, we find that active diffusion is driven by the actin cytoskeleton, although it is also enhanced by the flow. We also find that an important dynamic link exists between vesicles and cytoplasmic F-actin motion, as recently suggested in mouse oocytes.

Dynamics of epithelial monolayers has recently been interpreted in terms of a jamming or rigidity transition. How cells control such phase transitions is, however, unknown. Here we show that RAB5A, a key endocytic protein, is sufficient to induce large-scale, coordinated motility over tens of cells, and ballistic motion in otherwise kinetically arrested monolayers. This is linked to increased traction forces and to the extension of cell protrusions, which align with local velocity. Molecularly, impairing endocytosis, macropinocytosis or increasing fluid efflux abrogates RAB5A-induced collective motility. A simple model based on mechanical junctional tension and an active cell reorientation mechanism for the velocity of self-propelled cells identifies regimes of monolayer dynamics that explain endocytic reawakening of locomotion in terms of a combination of large-scale directed migration and local unjamming. These changes in multicellular dynamics enable collectives to migrate under physical constraints and may be exploited by tumours for interstitial dissemination.

We thank S Henkes for useful discussions. FGia and RC acknowledge funding from the Italian Ministry of University and Scientific Research (MIUR) under the program Futuro in Ricerca—Project ANISOFT (RBFR125H0M) and from Regione Lombardia and CARIPLO foundation under the joint action Avviso congiunto per l'incremento dell'attrattivita del sistema ricerca lombardo e della competitivita dei ricercatori candidati su strumenti ERC—Project 2016-0998. CM, SC and GS acknowledge funding from Associazione Italiana per la Ricerca sul Cancro (AIRC 10168 and 18621), MIUR, the Italian Ministry of Health, Ricerca Finalizzata (RF0235844), Worldwide Cancer Research (AICR-14-0335), and the European Research Council (Advanced-ERC-268836). CM was also supported by Fondazione Umberto Veronesi and SC by an AIRC fellowship. FGin acknowledges support from the Marie Curie Career Integration Grant (CIG) PCIG13-GA-2013-618399, and wish to thank the University of Milan and LibrOsteria for their hospitality while this work was underway.

Linearised fluctuating hydrodynamics describes effectively the concentration non-equilibrium fluctuations (NEF) arising during a diffusion process driven by a small concentration gradient. However, fluctuations in the presence of large gradients are not yet fully understood. Here we study the giant concentration NEF arising when a dense aqueous colloidal suspension is allowed to diffuse into an overlying layer of pure water. We use differential dynamic microscopy to determine both the statics and the dynamics of the fluctuations for several values of the wave-vector q. At small q, NEF are quenched by buoyancy, which prevents their full development and sets an upper timescale to their temporal relaxation. At intermediate q, the mean squared amplitude of NEF is characterised by a power law exponent -4, and fluctuations relax diffusively with diffusion coefficient D1. At large q, the amplitude of NEF vanishes and equilibrium concentration fluctuations are recovered, enabling a straightforward determination of the osmotic compressibility of the suspension during diffusion. In this q-range we also find that the relaxation of the fluctuations occurs with a diffusion coefficient D2 significantly different from D1. Both diffusion coefficients exhibit time-dependence with D1 increasing monotonically (by about 15%) and D2 showing the opposite behaviour (about 17% decrease). At equilibrium, the two coefficients coincide as expected. While the decrease of D2 is compatible with a diffusive evolution of the concentration profile, the increase of D1 is still not fully understood and may require considering nonlinearities that are neglected in current theories for highly stressed colloids.

We probe the roto-translational Brownian motion of optically anisotropic particles suspended in water with a simple and straightforward optical microscopy experiment that does not require positional or rotational particle tracking. We acquire a movie of the suspension placed between two polarizing elements and we extract the translational diffusion coefficient D T and the rotational diffusion coefficient D R from the analysis of the temporal correlation properties of the spatial Fourier modes of the intensity fluctuations in the movie. Our method is successfully tested with a dilute suspension of birefringent spherical colloidal particles obtained by polymerizing an emulsion of droplets of liquid crystal in a nematic phase, whose roto-translational dynamics is found to be well described by theory. The simplicity of our approach makes our method a viable alternative to particle tracking and depolarized dynamic light scattering.

The measurement of the intensity of light reflected by interfaces with extremely low reflectivity in water enables the label-free, multiplex detection of bio-molecular interactions. The binding of targets in solution to surface-immobilized probes (e.g. antibodies) can be quantified using a simple instrumental set-up, comprising a LED source and a camera. The condition of low reflectivity can be achieved by using a perfluorinated plastic substrate with refractive index very close to that of water. Here we show that, despite its simplicity, the method enables label-free detection in complex samples with high absorbance and turbidity. Diagnostic markers of Tomato spotted wilt virus are revealed in crude plant extracts of Datura stramonium leaves at early stage of infection. The measuring procedure requires only the addition of the sample into a cuvette and the acquisition of a series of images for a few minutes. The light reflectivity of the spots of specific antibodies increases more rapidly if the sample is prepared with infected plants.

When a macroscopic concentration gradient is present across a binary mixture, long-ranged non-equilibrium concentration fluctuations (NCF) appear as a consequence of the coupling between the gradient and spontaneous equilibrium velocity fluctuations. Long-ranged equilibrium concentration fluctuations (ECF) may be also observed when the mixture is close to a critical point. Here we study the interplay between NCF and critical ECF in a near critical mixture aniline/cyclohexane in the presence of a vertical concentration gradient. To this aim, we exploit a commercial optical microscope and a simple, custom-made, temperature-controlled cell to obtain simultaneous static and dynamic scattering information on the fluctuations. We first characterise the critical ECF at fixed temperature $T$ above the upper critical solution temperature $T_{c}$, in the wide temperature range $T-T_{c}\in[0.1,30]$ $^{o}$C. In this range, we observe the expected critical scaling behaviour for both the scattering intensity and the mass diffusion coefficient and we determine the critical exponents $\gamma$, $\nu$ and $\eta$, which are found in agreement with the 3D Ising values. We then study the system in the two-phase region ($TT_{i}$. During the transient, a vertical diffusive mass flux is present that causes the onset of NCF, whose amplitude vanishes with time, as the flux goes to zero. We also study the time dependence of the equilibrium scattering intensity $I_{eq}$, of the crossover wave-vector $q_{co}$ and of the diffusion coefficient $D$ during diffusion and find that all these quantities exhibit an exponential relaxation enslaved to the diffusive kinetics., Comment: 11 pages, 4 figures

Concentrated solutions of duplex-forming DNA oligomers organize into various mesophases among which is the nematic ( N ∗ ), which exhibits a macroscopic chiral helical precession of molecular orientation because of the chirality of the DNA molecule. Using a quantitative analysis of the transmission spectra in polarized optical microscopy, we have determined the handedness and pitch of this chiral nematic helix for a large number of sequences ranging from 8 to 20 bases. The B-DNA molecule exhibits a right-handed molecular double-helix structure that, for long molecules, always yields N ∗ phases with left-handed pitch in the μm range. We report here that ultrashort oligomeric duplexes show an extremely diverse behavior, with both left- and right-handed N ∗ helices and pitches ranging from macroscopic down to 0.3 μm. The behavior depends on the length and the sequence of the oligomers, and on the nature of the end-to-end interactions between helices. In particular, the N ∗ handedness strongly correlates with the oligomer length and concentration. Right-handed phases are found only for oligomers shorter than 14 base pairs, and for the sequences having the transition to the N ∗ phase at concentration larger than 620 mg/mL. Our findings indicate that in short DNA, the intermolecular double-helical interactions switch the preferred liquid crystal handedness when the columns of stacked duplexes are forced at high concentrations to separations comparable to the DNA double-helix pitch, a regime still to be theoretically described.