RESUMEN
Many natural materials display locally varying compositions that impart unique mechanical properties to them which are still unmatched by manmade counterparts. Synthetic materials often possess structures that are well-defined on the molecular level, but poorly defined on the microscale. A fundamental difference that leads to this dissimilarity between natural and synthetic materials is their processing. Many natural materials are assembled from compartmentalized reagents that are released in well-defined and spatially confined regions, resulting in locally varying compositions. By contrast, synthetic materials are typically processed in bulk. Inspired by nature, we introduce a drop-based technique that enables the design of microstructured hydrogel sheets possessing tuneable locally varying compositions. This control in the spatial composition and microstructure is achieved with a microfluidic Hele-Shaw cell that possesses traps with varying trapping strengths to selectively immobilize different types of drops. This modular platform is not limited to the fabrication of hydrogels but can be employed for any material that can be processed into drops and solidified within them. It likely opens up new possibilities for the design of structured, load-bearing hydrogels, as well as for the next generation of soft actuators and sensors.
RESUMEN
The microtubule cytoskeleton serves as a dynamic structural framework for mitosis in eukaryotic cells. TANGLED1 (TAN1) is a microtubule-binding protein that localizes to the division site and mitotic microtubules and plays a critical role in division plane orientation in plants. Here, in vitro experiments demonstrate that TAN1 directly binds microtubules, mediating microtubule zippering or end-on microtubule interactions, depending on their contact angle. Maize tan1 mutant cells improperly position the preprophase band (PPB), which predicts the future division site. However, cell shape-based modeling indicates that PPB positioning defects are likely a consequence of abnormal cell shapes and not due to TAN1 absence. In telophase, colocalization of growing microtubules ends from the phragmoplast with TAN1 at the division site suggests that TAN1 interacts with microtubule tips end-on. Together, our results suggest that TAN1 contributes to microtubule organization to ensure proper division plane orientation.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , División Celular , Microtúbulos/metabolismo , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Zea mays/metabolismo , Sitios de Unión , Proteínas de Ciclo Celular/genética , Regulación de la Expresión Génica de las Plantas , Microtúbulos/genética , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Unión Proteica , Transducción de Señal , Factores de Tiempo , Zea mays/genéticaRESUMEN
One key aspect of cell division in multicellular organisms is the orientation of the division plane. Proper division plane establishment contributes to normal plant body organization. To determine the importance of cell geometry in division plane orientation, we designed a three-dimensional probabilistic mathematical model to directly test the century-old hypothesis that cell divisions mimic soap-film minima. According to this hypothesis, daughter cells have equal volume and the division plane occurs where the surface area is at a minimum. We compared predicted division planes to a plant microtubule array that marks the division site, the preprophase band (PPB). PPB location typically matched one of the predicted divisions. Predicted divisions offset from the PPB occurred when a neighboring cell wall or PPB was directly adjacent to the predicted division site to avoid creating a potentially structurally unfavorable four-way junction. By comparing divisions of differently shaped plant cells (maize [Zea mays] epidermal cells and developing ligule cells and Arabidopsis thaliana guard cells) and animal cells (Caenorhabditis elegans embryonic cells) to divisions simulated in silico, we demonstrate the generality of this model to accurately predict in vivo division. This powerful model can be used to separate the contribution of geometry from mechanical stresses or developmental regulation in predicting division plane orientation.
Asunto(s)
Arabidopsis/citología , Modelos Biológicos , Células Vegetales/fisiología , Zea mays/citología , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Caenorhabditis elegans/citología , Caenorhabditis elegans/embriología , División Celular , Embrión no Mamífero/citología , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Microtúbulos/metabolismo , Microtúbulos/ultraestructura , Hojas de la Planta/citología , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Jabones/química , Imagen de Lapso de TiempoRESUMEN
Capillary open microsystems are attractive and increasingly used in biotechnology, biology, and diagnostics as they allow simple and reliable control of fluid flows. In contrast to closed microfluidic systems, however, two-phase capillary flows in open microfluidics have remained largely unexplored. In this work, we present the theoretical basis and experimental demonstration of a spontaneous capillary flow (SCF) of two-phase systems in open microchannels. Analytical results show that an immiscible plug placed in an open channel can never stop the SCF of a fluid in a uniform cross-section microchannel. Numerical investigations of the morphologies of immiscible plugs in a capillary flow reveal three different possible behaviors. Finally, the predicted behaviors of the plugs are demonstrated experimentally, revealing an effect of inertial forces on the plug behavior. A model for predicting plug behaviors in SCFs is proposed, enabling the design of open microfluidic droplet-based systems that are simple to fabricate and use. The open-channel approach to droplet-based microfluidics has the potential to enable applications in which each drop can be accessed at any time and any location with simple pipettes or other fluid dispensing systems.
RESUMEN
The microbial secondary metabolome encompasses great synthetic diversity, empowering microbes to tune their chemical responses to changing microenvironments. Traditional metabolomics methods are ill-equipped to probe a wide variety of environments or environmental dynamics. Here we introduce a class of microscale culture platforms to analyse chemical diversity of fungal and bacterial secondary metabolomes. By leveraging stable biphasic interfaces to integrate microculture with small molecule isolation via liquid-liquid extraction, we enable metabolomics-scale analysis using mass spectrometry. This platform facilitates exploration of culture microenvironments (including rare media typically inaccessible using established methods), unusual organic solvents for metabolite isolation and microbial mutants. Utilizing Aspergillus, a fungal genus known for its rich secondary metabolism, we characterize the effects of culture geometry and growth matrix on secondary metabolism, highlighting the potential use of microscale systems to unlock unknown or cryptic secondary metabolites for natural products discovery. Finally, we demonstrate the potential for this class of microfluidic systems to study interkingdom communication between fungi and bacteria.
Asunto(s)
Aspergillus/metabolismo , Fusarium/metabolismo , Metaboloma , Metabolómica/métodos , Pseudomonas aeruginosa/metabolismo , Ralstonia solanacearum/metabolismo , Aspergillus flavus/metabolismo , Aspergillus fumigatus/metabolismo , Aspergillus nidulans/metabolismo , Cromatografía Líquida de Alta Presión , Técnicas de Cocultivo , Técnicas de Cultivo , Microfluídica , Espectrometría de Masas en TándemRESUMEN
We study theoretically the influence of an external magnetic field on the orientation of an ellipsoidal magnetic particle adsorbed at a liquid interface. Using the finite element program Surface Evolver, we calculate the equilibrium meniscus shape around the ellipsoidal particle and its equilibrium tilt angle with respect to the undeformed interface θt when a magnetic field B is applied perpendicular to the interface. We find that as we increase field strength, θt increases and at a critical magnetic field Bc1 and tilt angle θc1, the particle undergoes a discontinuous transition to the 'perpendicular' orientation (θt = 90°). Our results agree qualitatively with the simplified theory of Bresme and Faraudo [F. Bresme and J. Faraudo, J. Phys.: Condens. Matter, 2007, 19, 375110] which assumes that the liquid interface is flat, while they agree quantitatively with recent lattice-Boltzmann simulations of Davies et al. [G. Davies et al., Soft Matter, 2014, 10, 6742] which account for the deformation of the liquid meniscus. We also show for the first time that upon reducing the external magnetic field, at a critical magnetic field Bc2 < Bc1, the particle undergoes a second discontinuous transition from the perpendicular orientation to a finite tilt angle θc2 < θc1. In other words, for micron-sized particles where the thermal energy kBT is negligible compared to the interfacial energy, the tilt angle vs. magnetic field curve exhibits hysteresis behaviour. Due to the higher degree of accuracy of the Surface Evolver method, we are able to analyse the behaviour of the particles near these orientational transitions accurately and study how the critical quantities Bc1, Bc2, θc1 and θc2 vary with particle aspect ratio and contact angle.
RESUMEN
Epithelial cells acquire functionally important shapes (e.g., squamous, cuboidal, columnar) during development. Here, we combine theory, quantitative imaging, and perturbations to analyze how tissue geometry, cell divisions, and mechanics interact to shape the presumptive enveloping layer (pre-EVL) on the zebrafish embryonic surface. We find that, under geometrical constraints, pre-EVL flattening is regulated by surface cell number changes following differentially oriented cell divisions. The division pattern is, in turn, determined by the cell shape distribution, which forms under geometrical constraints by cell-cell mechanical coupling. An integrated mathematical model of this shape-division feedback loop recapitulates empirical observations. Surprisingly, the model predicts that cell shape is robust to changes of tissue surface area, cell volume, and cell number, which we confirm in vivo. Further simulations and perturbations suggest the parameter linking cell shape and division orientation contributes to epithelial diversity. Together, our work identifies an evolvable design logic that enables robust cell-level regulation of tissue-level development.
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Células Epiteliales/citología , Modelos Biológicos , Morfogénesis , Pez Cebra/embriología , Animales , Fenómenos Biomecánicos , Recuento de Células , División Celular , Forma de la Célula , Embrión no Mamífero/citologíaRESUMEN
Inverse bicontinuous cubic lyotropic phases are a complex solution to the dilemma faced by all self-assembled water-amphiphile systems: how to satisfy the incompatible requirements for uniform interfacial curvature and uniform molecular packing. The solution reached in this case is for the water-amphiphile interfaces to deform hyperbolically onto triply periodic minimal surfaces. We have previously suggested that although the molecular packing in these structures is rather uniform the relative phase behavior of the gyroid, double diamond, and primitive inverse bicontinuous cubic phases can be understood in terms of subtle differences in packing frustration. In this work, we have calculated the packing frustration for these cubics under the constraint that their interfaces have constant mean curvature. We find that the relative packing stress does indeed differ between phases. The gyroid cubic has the least packing stress, and at low water volume fraction, the primitive cubic has the greatest packing stress. However, at very high water volume fraction, the double diamond cubic becomes the structure with the greatest packing stress. We have tested the model in two ways. For a system with a double diamond cubic phase in excess water, the addition of a hydrophobe may release packing frustration and preferentially stabilize the primitive cubic, since this has previously been shown to have lower curvature elastic energy. We have confirmed this prediction by adding the long chain alkane tricosane to 1-monoolein in excess water. The model also predicts that if one were able to hydrate the double diamond cubic to high water volume fractions, one should destabilize the phase with respect to the primitive cubic. We have found that such highly swollen metastable bicontinuous cubic phases can be formed within onion vesicles. Data from monoelaidin in excess water display a well-defined transition, with the primitive cubic appearing above a water volume fraction of 0.75. Both of these results lend support to the proposition that differences in the packing frustration between inverse bicontinuous cubic phases play a pivotal role in their relative phase stability.