During embryo development, patterns of protein concentration appear in response to morphogen gradients. These patterns provide spatial and chemical information that directs the fate of the underlying cells. Here, we emulate this process within non-living matter and demonstrate the autonomous structuration of a synthetic material. First, we use DNA-based reaction networks to synthesize a French flag, an archetypal pattern composed of three chemically distinct zones with sharp borders whose synthetic analogue has remained elusive. A bistable network within a shallow concentration gradient creates an immobile, sharp and long-lasting concentration front through a reaction-diffusion mechanism. The combination of two bistable circuits generates a French flag pattern whose 'phenotype' can be reprogrammed by network mutation. Second, these concentration patterns control the macroscopic organization of DNA-decorated particles, inducing a French flag pattern of colloidal aggregation. This experimental framework could be used to test reaction-diffusion models and fabricate soft materials following an autonomous developmental programme.
We introduce a DNA-based reaction-diffusion (RD) system in which reaction and diffusion terms can be precisely and independently controlled. The effective diffusion coefficient of an individual reaction component, as we demonstrate on a traveling wave, can be reduced up to 2.7-fold using a self-assembled hydrodynamic drag. The intrinsic programmability of this RD system allows us to engineer, for the first time, orthogonal autocatalysts that counterpropagate with minimal interaction. Our results are in excellent quantitative agreement with predictions of the Fisher-Kolmogorov-Petrovskii-Piscunov model. These advances open the way for the rational engineering of pattern formation in pure chemical RD systems.
Multilayer soft lithography of polydimethylsiloxane (PDMS) is a well-known method for the fabrication of complex fluidic functions. With advantages and drawbacks, this technique allows fabrication of valves, pumps and micro-mixers. However, the process is inadequate for industrial applications. Here, we report a rapid prototyping technique for the fabrication of multilayer microfluidic devices, using a different and promising class of polymers. Using styrenic thermoplastic elastomers (TPE), we demonstrate a rapid technique for the fabrication and assembly of pneumatically driven valves in a multilayer microfluidic device made completely from thermoplastics. This material solution is transparent, biocompatible and as flexible as PDMS, and has high throughput thermoforming processing characteristics. We established a proof of principle for valving and mixing with three different grades of TPE using an SU-8 master mold. Specific viscoelastic properties of each grade allow us to report enhanced bonding capabilities from room temperature bonding to free pressure thermally assisted bonding. In terms of microfabrication, beyond classically embossing means, we demonstrate a high-throughput thermoforming method, where TPE molding experiments have been carried out without applied pressure and vacuum assistance within an overall cycle time of 180 s. The quality of the obtained thermoplastic systems show robust behavior and an opening/closing frequency of 5 Hz.
Active matter locally converts chemical energy into mechanical work and, for this reason, it provides new mechanisms of pattern formation. In particular, active gels made of protein motors and filaments are far-from-equilibrium systems that exhibit spontaneous flow, 1,2 leading to active turbulence in two and three dimensions 3,4 and coherent flow in three dimensions 5 (3D). Although these dynamic flows reveal a characteristic length scale resulting from the interplay between active forcing and passive restoring forces, the observation of static and long-range spatial patterns in active gels has remained elusive. In this work, we demonstrate that a 2D free-standing nematic active gel, formed spontaneously by depletion forces from a 3D solution of kinesin motors and microtubule filaments, actively buckles out-of-plane into a centimeter-sized periodic corrugated sheet that is stable for several days at low activity. Importantly, the corrugations are formed in the absence of flow and their wavelength and stability are controlled by the motor concentration, in agreement with a hydrodynamic theory.At higher activities these patterns are transient with the gel becoming turbulent at longer times. Our results underline the importance of both passive and active forces in shaping active gels and indicate that a static material can be sculpted through an active mechanism.
We report on results of design and fabrication of a microfluidic dye laser that consists of a ring resonator, a waveguide for laser emission output, and microfluidic elements for flow control, all integrated on a chip. The optical resonator and the waveguide were obtained by photolithography, whereas microfluidic elements such as channels, valves, and pumps were fabricated by multilayer soft lithography. As results, the prototype device worked with a few nanoliters of Rhodamine 6G dye molecules in ethanol solution and showed a laser threshold of ∼15μJ∕mm2 when optically pumped with a frequency doubled Nd:YAG laser at 532nm wavelength. The modification of the laser output intensity due to photobleaching effect was characterized by changing the dye flow velocity through the cavity. In addition, the emission wavelength of the laser could be easily tuned by changing the dye molecule concentration with the integrated microfluidic elements.
This article describes the fabrication of microfluidic networks (lFNs) from a commercially available (styrene)-(ethylene/butylene)-(styrene) (SEBS) block copolymer (BCP). The unique combination of hard and elastomeric properties provided by this material promotes high-throughput replication of fluidic structures using thermoforming technologies, while retaining the advantage of quick and easy assembly via conformal contact, as commonly achieved for devices fabricated from poly(dimethylsiloxane (PDMS). We employ Versaflex CL30, which is optically transparent, available at low cost (e.g., $2.50/Lbs), and likely to be compatible with a broad range of biological species. We demonstrate excellent fidelity in replication of fluidic structures using hot embossing lithography in conjunction with a photolithographically prepared Si/SU-8 master mold. Moreover, we introduce rapid prototyping of high-quality structures using an approach that we call soft thermoplastic lithography (STPL). Thanks to the rheological characteristics of the SEBS copolymer, STPL enables thermoforming on a heated master at temperatures around 170°C. Using this approach, replication can be completed within a very short period of time (e.g., less than 3 min) without the need of resorting to pressure-or vacuum-assisted instrumentation. Serving as a proof-of-concept, we devise a lFN that is suitable for the formation of miniaturized arrays comprising fluorescently labeled oligonucleotides and proteins on hard plastic substrates. Resultant spots are characterized by high fluorescent contrast, excellent edge definition, and uniform distribution of probes within the modified areas.
In this paper we first introduce a novel fabrication process, which allows for easy integration of thin track-etched nanoporous membranes, within 2D or 3D microchannel networks. In these networks, soluble chemical compounds can diffuse out of the channels through well-defined and spatially organized microfabricated porous openings. Interestingly, multiple micron-scale porous areas can be integrated in the same device and each of these areas can be connected to a different microfluidic channel and reservoir. We then present and characterize several membrane-based microdevices and their use for the generation of stable diffusible concentration gradients and complex dynamic chemical landscapes under shear free conditions. We also demonstrate how a simple flow-focusing geometry can be used to generate "on-demand" concentration profiles. In turn, these devices should provide an ideal experimental framework for high throughput cell-based assays: long term high-resolution video microscopy experiments can be performed, under multiple spatially and temporally controlled chemical conditions, with simple protocols and in a cell-friendly environment.
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