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We present an efficient method for investigating kinetics of gelling system, demonstrating that combining active learning and microrheology can streamline response surface construction and predict how gelation times influence the cell morphology. 

Abstract During morphogenesis, cells collectively execute directional forces that drive the programmed folding and growth of the layers, forming tissues and organs. The ability to recapitulate aspects of these processes in  vitro will constitute a significant leap forward in the field of tissue engineering. Free‐standing, self‐organizing, cell‐laden matrices are fabricated using a sequential deposition approach that uses liquid crystal‐templated hydrogel fibers to direct cell arrangements. The orientation of hydrogel fibers is controlled using flow or boundary cues, while their microstructures are controlled by depletion interaction and probed by scattering and microscopy. These fibers effectively direct cells embedded in a collagen matrix, creating multilayer structures through contact guidance and by leveraging steric interactions amongst the cells. In uniformly aligned cell matrices, oriented cells exert traction forces that can induce preferential contraction of the matrix. Simultaneously, the matrix densifies and develops anisotropy through cell remodeling. Such an approach can be extended to create cell arrangements with arbitrary in‐plane patterns, allowing for coordinated cell forces and pre‐programmed, macroscopic shape changes. This work reveals a fundamentally new path for controlled force generation, emphasizing the role of a carefully designed initial orientational field for manipulating shape transformations of reconstituted matrices. 

High-throughput microrheology and simple viscosity modeling can be used to continuously monitor the kinetic evolution of polymer molecular weight during controlled polymerizations. 

The ability of cells to reorganize in response to external stimuli is important in areas ranging from morphogenesis to tissue engineering. While nematic order is common in biological tissues, it typically only extends to small regions of cells interacting via steric repulsion. On isotropic substrates, elongated cells can co-align due to steric effects, forming ordered but randomly oriented finite-size domains. However, we have discovered that flat substrates with nematic order can induce global nematic alignment of dense, spindle-like cells, thereby influencing cell organization and collective motion and driving alignment on the scale of the entire tissue. Remarkably, single cells are not sensitive to the substrate’s anisotropy. Rather, the emergence of global nematic order is a collective phenomenon that requires both steric effects and molecular-scale anisotropy of the substrate. To quantify the rich set of behaviours afforded by this system, we analyse velocity, positional and orientational correlations for several thousand cells over days. The establishment of global order is facilitated by enhanced cell division along the substrate’s nematic axis, and associated extensile stresses that restructure the cells’ actomyosin networks. Our work provides a new understanding of the dynamics of cellular remodelling and organization among weakly interacting cells. 

Evolution of composition, rheology, and morphology during phase separation in complex fluids is highly coupled to rheological and mass transport processes within the emerging phases, and understanding this coupling is critical for materials design of multiphase complex fluids. Characterizing these dependencies typically requires careful measurement of a large number of equilibrium and transport properties that are difficult to measure in situ as phase separation proceeds. Here, we propose and demonstrate a high-throughput microscopy platform to achieve simultaneous, in situ mapping of time-evolving morphology and microrheology in phase separating complex fluids over a large compositional space. The method was applied to a canonical example of polyelectrolyte complex coacervation, whereby mixing of oppositely charged species leads to liquid–liquid phase separation into distinct solute-dense and dilute phases. Morphology and rheology were measured simultaneously and kinetically after mixing to track the progression of phase separation. Once equilibrated, the dense phase viscosity was determined to high compositional accuracy using passive probe microrheology, and the results were used to derive empirical relationships between the composition and viscosity. These relationships were inverted to reconstruct the dense phase boundary itself, and further extended to other mixture compositions. The resulting predictions were validated by independent equilibrium compositional measurements. This platform paves the way for rapid screening and formulation of complex fluids and (bio)macromolecular materials, and serves as a critical link between formulation and rheology for multi-phase material discovery.