A Venturi microregulator array module for distributed pressure control.

Pressure-driven flow control systems are a critical component in many microfluidic devices. Compartmentalization of this functionality into a stand-alone module possessing a simple interface would allow reduction of the number of pneumatic interconnects required for fluidic control. Ideally, such a module would also be sufficiently compact for implementation in portable platforms. In our current work, we show the feasibility of using a modular array of Venturi pressure microregulators for coordinated droplet manipulation. The arrayed microregulators share a single pressure input and are capable of outputting electronically controlled pressures that can be independently set between ±1.3 kPa. Because the Venturi microregulator operates by thermal perturbation of a choked gas flow, this output range corresponds to a temperature variation between 20 and 95°C. Using the array, we demonstrate loading, splitting, merging, and independent movement of multiple droplets in a valveless microchannel network.

Selective arraying of complex particle patterns.

Procedures requiring precise and accurate positioning of particles and cells have impacted a broad range of research interests including molecular detection, self-assembly and tissue and cell engineering. These fields would be greatly aided by more advanced, yet straightforward, micro-object positioning methods that are precise, scalable, responsive and flexible. We have developed an arrayed, multilayer surface patterned microfluidic device which uses laminar convective flow to actively position particles into any desired, two-dimensional, predesigned pattern. Objects including 10 microm polystyrene particles and Saccharomycodes ludwigii cells are rapidly (approximately 2 s) loaded onto vacuum-actuated holes, allowing us to both generate anisotropic particles and culture S. ludwigii cells. The device was further modified to individually control two sets of holes, adding control of pattern composition. With rapid, precise and adaptable operation, multilayer microfluidic devices should greatly assist in research where precise object placement and proximity is necessary.

Acoustically driven programmable liquid motion using resonance cavities.

Performance and utility of microfluidic systems are often overshadowed by the difficulties and costs associated with operation and control. As a step toward the development of a more efficient platform for microfluidic control, we present a distributed pressure generation scheme whereby independently tunable pressure sources can be simultaneously controlled by using a single acoustic source. We demonstrate how this scheme can be used to perform precise droplet positioning as well as merging, splitting, and sorting within open microfluidic networks. We further show how this scheme can be implemented for control of continuous-flow systems, specifically for generation of acoustically tunable liquid gradients. Device operation hinges on a resonance-decoding and rectification mechanism by which the frequency content in a composite acoustic input is decomposed into multiple independently buffered output pressures. The device consists of a bank of 4 uniquely tuned resonance cavities (404, 484, 532, and 654 Hz), each being responsible for the actuation of a single droplet, 4 identical flow-rectification structures, and a single acoustic source. Cavities selectively amplify resonant tones in the input signal, resulting in highly elevated local cavity pressures. Fluidic-rectification structures then serve to convert the elevated oscillating cavity pressures into unidirectional flows. The resulting pressure gradients, which are used to manipulate fluids in a microdevice, are tunable over a range of approximately 0-200 Pa with a control resolution of 10 Pa.

An electronic Venturi-based pressure microregulator.

Microfluidic systems often use pressure-driven flow to induce fluidic motion, but control of pumps and valves can necessitate numerous external connections or an extensive external control infrastructure. Here, we describe an electronically controlled pressure microregulator that can output pressures both greater and less than atmospheric pressure over a range of 2 kPa from a single pressurized air input of 110 kPa. Multiple independently controlled microregulators integrated in one device can potentially share the same air input. The microregulator operates by using embedded resistive heaters to vary the temperature of a gas flowing through a converging-diverging Venturi nozzle between 25 degrees C and 85 degrees C with a resolution of 33 Pa degrees C(-1). We established the switching speed of the microregulator by accurately moving 1 microL droplets of water in a microchannel via pneumatic propulsion. Droplet deceleration from approximately 1 cm s(-1) to zero velocity required less than 0.8 s. The component is readily integrable into most device designs containing fluidic channels and electronics without introducing additional fabrication complexity.

Endothelial cell diversity revealed by global expression profiling.

The vascular system is locally specialized to accommodate widely varying blood flow and pressure and the distinct needs of individual tissues. The endothelial cells (ECs) that line the lumens of blood and lymphatic vessels play an integral role in the regional specialization of vascular structure and physiology. However, our understanding of EC diversity is limited. To explore EC specialization on a global scale, we used DNA microarrays to determine the expression profile of 53 cultured ECs. We found that ECs from different blood vessels and microvascular ECs from different tissues have distinct and characteristic gene expression profiles. Pervasive differences in gene expression patterns distinguish the ECs of large vessels from microvascular ECs. We identified groups of genes characteristic of arterial and venous endothelium. Hey2, the human homologue of the zebrafish gene gridlock, was selectively expressed in arterial ECs and induced the expression of several arterial-specific genes. Several genes critical in the establishment of left/right asymmetry were expressed preferentially in venous ECs, suggesting coordination between vascular differentiation and body plan development. Tissue-specific expression patterns in different tissue microvascular ECs suggest they are distinct differentiated cell types that play roles in the local physiology of their respective organs and tissues.

Genomewide view of gene silencing by small interfering RNAs.

RNA interference (RNAi) is an evolutionarily conserved mechanism in plant and animal cells that directs the degradation of messenger RNAs homologous to short double-stranded RNAs termed small interfering RNA (siRNA). The ability of siRNA to direct gene silencing in mammalian cells has raised the possibility that siRNA might be used to investigate gene function in a high throughput fashion or to modulate gene expression in human diseases. The specificity of siRNA-mediated silencing, a critical consideration in these applications, has not been addressed on a genomewide scale. Here we show that siRNA-induced gene silencing of transient or stably expressed mRNA is highly gene-specific and does not produce secondary effects detectable by genomewide expression profiling. A test for transitive RNAi, extension of the RNAi effect to sequences 5' of the target region that has been observed in Caenorhabditis elegans, was unable to detect this phenomenon in human cells.