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World-to-chip (macro-to-micro) interface continues to be one of the most complicated, ineffective, and unreliable components in the development of emerging lab-on-a-chip systems involving integrated microfluidic operations. A number of irreversible (e.g., adhesive gluing) and reversible techniques (e.g., press fitting) have attempted to provide dedicated fluidic passage from standard tubing to miniature on-chip devices, none of which completely addresses the above concerns. In this paper, we present standardized adhesive-free microfluidic adaptors, referred to as Fit-to-Flow (F2F) Interconnects, to achieve reliable hermetic seal, high-density tube packing, self-aligned plug-in, reworkable connectivity, straightforward scalability and expandability, and applicability to broad lab-on-a-chip platforms; analogous to the modular plug-and-play USB architecture employed in modern electronics. Specifically, two distinct physical packaging mechanisms are applied, with one utilizing induced tensile stress in elastomeric socket to establish reversible seal and the other using negative pressure to provide on demand vacuum shield, both of which can be adapted to a variety of experimental configurations. The non-leaking performance (up to 336 kPa) along with high tube-packing density (of 1 tube/mm2) and accurate self-guided alignment (of 10 μm) have been characterized. In addition, a 3D microfluidic mixer and a 6-level chemical gradient generator paired with the corresponding F2F Interconnects have been devised to illustrate the applicability of the universal fluidic connections to classic lab-on-a-chip operations. [ABSTRACT FROM AUTHOR]

As an emerging alternative to the conventional counterpart, surface microfluidics incorporates both intrinsic resistive solid-liquid and elastic frictionless gas-liquid interfaces, leading to unique flow-pressure characteristics. Furthermore, the open-surface microfluidic platforms can be fabricated on a monolithic substrate with high wettability contrast by the previously reported one-step lithographic process of a photosensitive superhydrophobic nanocomposite material, which permits flexible fluidic operations and direct surface modifications. In the paper, we first present three-dimensional microfluidic manipulations utilizing the unconventional gas-liquid interfaces of surface microfluidics, outlined by the micropatterned wetting boundaries (also known as the triple lines). In contrast to the primary linear (resistive) nature of the conventional closed-channel microfluidics, the distinct elastic interface of surface microfluidics enables remarkable three-dimensional (deformable) and time-dependent (capacitive) operations of the flow. Specifically, spatiotemporal dependence of microflow patterns on the planar fluidic surfaces has been theoretically analyzed and experimentally characterized. Utilizing the unconventional interface-enabled flow-pressure relationship, novel surface fluidic operations, including microflow regulation and flow-controlled switching, have been demonstrated and fully investigated. Furthermore, three-dimensional surface microfluidic networks together with analog-to-digital stereo-flow activations have been established, in which miniature capillary bridges form fluidic connections between two independent surface microfluidic circuits. [ABSTRACT FROM AUTHOR]

In this paper, we first report on directphotolithography- based microfabrication of transparent superhydrophobic micropatterns using novel photodefinable nanocomposites, combining the nanomorphology and hydrophobicity of polytetrafluoroethylene (PTFE) nanoparticles and the photopatternability and transparency of an SU-8 photoresist using both direct-mixing and coating-immobilization methods. The direct mixture of PTFE-SU-8 nanocomposite can be reliably spin-coated and photopatterned onto transparent substrates (e.g., glass or polymers) with a minimal feature resolution of 50 μm. The resulting nanocomposite film possesses a contact angle of water at 150°, although its optical transparency is less than 30%. Furthermore, a modified coating-immobilization approach, employing spray coating and thermal immobilization of PTFE nanoparticles onto an SU-8 polymer matrix, significantly enhances superhydrophobicity, lithography resolution, as well as optical transparency. The highest optical transparency of 80% and a minimal feature resolution of 10 μm have been achieved using the standard photolithography approach, while the contact angle of water above 165° enables extraordinary superhydrophobicity with low hysteresis. The novel PTFE-SU-8 nanocomposites provide a unique combination of superhydrophobicity, optical transparency, and photopatternability, along with excellent adaptability and simple processability, which offer great extension to rapidly evolving micro-nanoengineering applications. [ABSTRACT FROM AUTHOR]

In this paper, we present a simple adhesive-free interconnect technology for microfluidic systems. Reliable connection between microfluidic devices and macroscopic world is a critical issue in many lab-on-a-chip applications. In such interconnections, it is particularly advantageous to avoid liquid adhesives (e.g., epoxy, silicone, etc.) since they can create reliability (blockage or leakage) and reproducibility problems. Our adhesive-free interconnection technology is based on a micromachined silicon flange which is connected to a medical-grade silastic tube using a heat shrink tubing sleeve. The silicon part was fabricated by consecutive anisotropic and isotropic etching processes resulting in a through-hole surrounded by a connection socket ring created using the well known lag effect of deep reactive ion etching. Bio- compatible polyolelin heat shrink tubing (expansion ratio of 2:1 at its shrink temperature of 143 °C) was used to connect and tightly seal the silicon flange to the silastic tube without application of any adhesive. A two-step heat treatment starting from the silicon flange and terminating at the silastic tubing was used to seal the connection. High temperature removability of the heat shrink tubing makes interconnects reworkable. Pull-out force was measured to test the mechanical strength of interconnects with different socket shapes (square, hexagon, octagon, and 32-side circle). Increasing the number of sides improves the mechanical strength with the circular interconnects having an average pull-out force of more than 3N. Leakage tests were also performed to characterize the seal quality. The circular connectors of two different flange sizes showed zero leakage up to the maximum test pressure of 200 kPa (29 psi). [ABSTRACT FROM AUTHOR]

Abstract: Intracellular ice formation (IIF) plays a crucial role in cryobiology. The cell membrane is believed to play important roles in IIF initiation, however the underlying biophysical mechanisms are still not entirely understood. In this work we use a unidirectional freezing design for observations of the freezing process across the cell membrane, which then allow cell membrane integrity detection via dual fluorescent staining. Important IIF information including the location of the initiation site, the growth rate, and the ice dendrite morphology was well obtained. It is found that IIF always initiates at the cell membrane in contact with extracellular ice. Cell membranes are found to be disrupted after IIF in the high freezing rate region (>20°C/min), however they are observed to keep their integrity after IIF in the low freezing rate region (<20°C/min). The extracellular to intracellular ice penetration seems to be easier and the ice dendrites become larger when IIF is applied again to the intact cells in subsequent freezing cycles. The results give insight into the IIF mechanisms as well as the relationship between IIF and the cell membrane. [Copyright &y& Elsevier]

Droplet-based transport phenomena driven by surface tension have been explored as an automated pumping source for a number of chemical and biological applications. In this paper, we present a comprehensive theoretical and experimental investigation of unconventional droplet-based motions on a superhydrophobic-patterned surface microfluidic (S2M) platform. The S2M surfaces are monolithically fabricated using a facile two-step laser micromachining technique on regular polydimethylsiloxane (PDMS) chemistry. Unlike the traditional droplet-driven pumps built on an enclosed microfluidic network, the S2M network pins the liquid–solid interface of droplets to the lithographically defined wetting boundary and establishes a direct linkage between the volumetric and hydraulic measures. Moreover, diverse modes of droplet motions are theoretically determined and experimentally characterized in a bi-droplet configuration, among which several unconventional droplet-driven transport phenomena are first demonstrated. These include big-to-small droplet merging, droplet balancing, as well as bidirectional transporting, in addition to the classic small-to-big droplet transition. Furthermore, multi-stage programmable bidirectional pumping has been implemented on the S2M platform, according to the newly established droplet manipulation principle, to illustrate its potential use for automated biomicrofluidic and point-of-care diagnostic applications. [ABSTRACT FROM AUTHOR]

In this paper, we present a novel rapid-prototyping process for out-of-cleanroom microfabrication of three-dimensional multilayer microfluidic structures with a 10 µm resolution, referred to as the Direct Projection on Dry-film Photoresist (DP2). A commercially available digital projector is customized to function as a direct mask generation and photo exposure system, while easy-processing photosensitive dry films are used as the microfluidic constructs. Multilayer alignments among maskless-patterned layers are reliably achieved by using a Software Alignment technique with less than 10 µm precision, which eliminates the use of mechanical travelling stage. The bonding between different layers of dry film, simply enabled by a plasma-assisted thermal lamination, offers an easy implementation for suspended multilayer microstructures. Development of a complex microfluidic chip from computer layout can thus be accomplished within an hour in a regular chemical or biological lab environment using this approach. [ABSTRACT FROM AUTHOR]

With the major advances in soft lithography and polymer materials, use of microfluidic devices has attracted tremendous attention recently. A simple and fast micromachining process is highly in demand to prototype such a device efficiently and economically. In this paper, we first reported an out-of-cleanroom printing-based integrated microfabrication process, referred to as the lab-on-a-print(LOP), for rapid-prototyping three-dimensional microfluidics. Using this lab-on-a-printprocess, we demonstrated the potential to accomplish an entire design-to-fabrication cycle within an hour, including about 70 µm resolution of direct-lithography patterning, well-controlled polyimide wet etching, three-dimensional pattern alignment and multilayer wax thermal-fusion packaging. A microfluidic gradient generator was prepared and tested for validation of the lab-on-a-printmicrofabrication process. [ABSTRACT FROM AUTHOR]

In this paper, we introduce a novel microfluidic combinatorial synthesis platform, referred to as Microfluidic Print-to-Synthesis (MPS), for custom high-throughput and automated synthesis of a large number of unique peptides in a microarray format. The MPS method utilizes standard Fmoc chemistry to link amino acids on a polyethylene glycol (PEG)-functionalized microdisc array. The resulting peptide microarrays permit rapid screening for interactions with molecular targets or live cells, with low nonspecific binding. Such combinatorial peptide microarrays can be reliably prepared at a spot size of 200 µm with 1 mm center-tocenter distance, dimensions that require only minimal reagent consumption (less than 30 nL per spot per coupling reaction). The MPS platform has a scalable design for extended multiplexibility, allowing for 12 different building blocks and coupling reagents to be dispensed in one microfluidic cartridge in the current format, and could be further scaled up. As proof of concept for the MPS platform, we designed and constructed a focused tetrapeptide library featuring 2560 synthetic peptide sequences, capped at the N-terminus with 4-[(N'-2-methylphenyl)ureido]phenylacetic acid. We then used live human T lymphocyte Jurkat cells as a probe to screen the peptide microarrays for their interaction with a4ß1 integrin overexpressed and activated on these cells. Unlike the one-bead-one-compound approach that requires subsequent decoding of positive beads, each spot in the MPS array is spatially addressable. Therefore, this platform is an ideal tool for rapid optimization of lead compounds found in nature or discovered from diverse combinatorial libraries, using either biochemical or cell-based assays. [ABSTRACT FROM AUTHOR]

Since the 1960s, combination chemotherapy has been widely utilized as a standard method to treat cancer. However, because of the potentially enormous number of drug candidates and combinations, conventional identification methods of the effective drug combinations are usually associated with significantly high operational costs, low throughput screening, laborious and time-consuming procedures, and ethical concerns. In this paper, we present a low-cost, high-efficiency microfluidic print-to-screen (P2S) platform, which integrates combinatorial screening with biomolecular printing for high-throughput screening of anticancer drug combinations. This P2S platform provides several distinct advantages and features, including automatic combinatorial printing, high-throughput parallel drug screening, modular disposable cartridge, and biocompatibility, which can potentially speed up the entire discovery cycle of potent drug combinations. Microfluidic impact printing utilizing plug-and-play microfluidic cartridges is experimentally characterized with controllable droplet volume and accurate positioning. Furthermore, the combinatorial print-to-screen assay is demonstrated in a proof-of-concept biological experiment which can identify the positive hits among the entire drug combination library in a parallel and rapid manner. Overall, this microfluidic print-to-screen platform offers a simple, low-cost, high-efficiency solution for high-throughput large-scale combinatorial screening and can be applicable for various emerging applications in drug cocktail discovery. [ABSTRACT FROM AUTHOR]

Microfluidic impact printing has been recently introduced, utilizing its nature of simple device architecture, low cost, non-contamination, and scalable multiplexability and high throughput. In this paper, we have introduced an impact-based droplet printing platform utilizing a simple plug-and-play microfluidic cartridge driven by piezoelectric actuators. Such a customizable printing system allows for ultrafine control of droplet volume from picoliters (23 pl) to nanoliters (10 nl), a 500 fold variation. The high flexibility of droplet generation can be simply achieved by controlling the magnitude of actuation (e.g., driving voltage) and the waveform shape of actuation pulses, in addition to nozzle size restrictions. Detailed printing characterizations on these parameters have been conducted consecutively. A multiplexed impact printing system has been prototyped and demonstrated to provide the functions of single-droplet jetting and droplet multiplexing as well as concentration gradient generation. Moreover, a generic biological assay has also been tested and validated on this printing platform. Therefore, the microfluidic droplet printing system could be of potential value to establish multiplexed micro reactors for high-throughput life science applications. [ABSTRACT FROM AUTHOR]

The role that posterior parietal (PPC) and motor cortices play in modulating neural responses in somatosensory areas 1 and 2 was examined with reversible deactivation by transient cooling. Multiunit recordings from neurons in areas 1 and 2 were collected from six anesthetized adult monkeys (Macaca mulatta) before, during, and after reversible deactivation of areas 5L or 7b or motor cortex (M1/PM), while select locations on the hand and forelimb were stimulated. Response changes were quantified as increases and decreases to stimulus-driven activity relative to baseline and analyzed during three recording epochs: during deactivation ("cool") and at two time points after deactivation ("rewarm 1," "rewarm 2"). Although the type of response change observed was variable, for neurons at the recording sites tested >90% exhibited a significant change in response during cooling of 7b while cooling area 5L or M1/PM produced a change in 75% and 64% of sites, respectively. These results suggest that regions in the PPC, and to a lesser extent motor cortex, shape the response characteristics of neurons in areas 1 and 2 and that this kind of feedback modulation is necessary for normal somatosensory processing. Furthermore, this modulation appears to happen on a minute-by-minute basis and may serve as the substrate for phenomena such as somatosensory attention. [ABSTRACT FROM AUTHOR]

Somatosensory processing in the anesthetized macaque monkey was examined by reversibly deactivating posterior parietal areas 5L and 7b and motor/premotor cortex (M1/ PM) with microfluidic thermal regulators developed by our laboratories. We examined changes in receptive field size and configuration for neurons in areas 1 and 2 that occurred during and after cooling deactivation. Together the deactivated fields and areas 1 and 2 form part of a network for reaching and grasping in human and nonhuman primates. Cooling area 7b had a dramatic effect on receptive field size for neurons in areas 1 and 2, while cooling area 5 had moderate effects and cooling M1/PM had little effect. Specifically, cooling discrete locations in 7b resulted in expansions of the receptive fields for neurons in areas 1 and 2 that were greater in magnitude and occurred in a higher proportion of sites than similar changes evoked by cooling the other fields. At some sites, the neural receptive field returned to the precooling configuration within 5-22 min of rewarming, but at other sites changes in receptive fields persisted. These results indicate that there are profound top-down influences on sensory processing of early cortical areas in the somatosensory cortex. [ABSTRACT FROM AUTHOR]

The emerging technologies on mobile-based diagnosis and bioanalytical detection have enabled powerful laboratory assays such as enzyme-linked immunosorbent assay (ELISA) to be conducted in field-use lab-on-a-chip devices. In this paper, we present a low-cost universal serial bus (USB)-interfaced mobile platform to perform microfluidic ELISA operations in detecting the presence and concentrations of BDE-47 (2,2',4,4'-tetrabromodiphenyl ether), an environmental contaminant found in our food supply with adverse health impact. Our point-of-care diagnostic device utilizes flexible interdigitated carbon black electrodes to convert electric current into a microfluidic pump via gas bubble expansion during electrolytic reaction. The micropump receives power from a mobile phone and transports BDE-47 analytes through the microfluidic device conducting competitive ELISA. Using variable domain of heavy chain antibodies (commonly referred to as single domain antibodies or Nanobodies), the proposed device is sensitive for a BDE-47 concentration range of 10-3-104 µg/l, with a comparable performance to that uses a standard competitive ELISA protocol. It is anticipated that the potential impact in mobile detection of health and environmental contaminants will prove beneficial to our community and low-resource environments. [ABSTRACT FROM AUTHOR]

In this paper, we report on the effect of patterned non-adhesive hydrogel nanosurfaces on the self-assembly of highly ordered colloids. Polyethylene glycol (PEG) hydrogel is employed as the substrate material in the study, for its desired non-adhesive property, and biocompatibility as well as photopatternability. Ultrafine PEG features are photopatterned onto glass substrates with minimal feature resolution of 500 nm using ultraviolet or deep ultraviolet exposure. By simply controlling the colloidal concentration of the nanoassembly solutions and the dimensions of the wells, a range of highly organized nanocolloidal patterns are formed inside the PEG wells. Unlike the traditional surface modification techniques, ours takes advantage of the unique non-adhesive property of PEG hydrogels to achieve extremely high selectivity in the pattern-assisted nanoassembly. Our experiments show that with oxygen plasma treatment, the non-adhesive property of the PEG surface deteriorates significantly, leading to non-selective assembly with complete surface coverage of nanocolloidal beads under the same processing condition. Therefore, benefiting from the unique non-adhesive surface property, the pattern-assisted nanoassembly method enables a highly predictable and robust process for colloidal nanofabrication, and the obtained nanocolloidal arrays with well organized patterns could potentially find applications in photonic crystal fabrication, biological sensing and analytical detection. [ABSTRACT FROM AUTHOR]

We have developed a compact and lightweight microfluidic cooling device to reversibly deactivate one or more areas of the neocortex to examine its functional macrocircuitry as well as behavioral and cortical plasticity. The device, which we term the "cooling chip," consists of thin silicone tubing (through which chilled ethanol is circulated) embedded in mechanically compliant polydimethylsiloxane (PDMS). PDMS is tailored to compact device dimensions (as small as 21 mm3) that precisely accommodate the geometry of the targeted cortical area. The biocompatible design makes it suitable for both acute preparations and chronic implantation for long-term behavioral studies. The cooling chip accommodates an in-cortex microthermocouple measuring local cortical temperature. A microelectrode may be used to record simultaneous neural responses at the same location. Cortex temperature is controlled by computer regulation of the coolant flow, which can achieve a localized cortical temperature drop from 37 to 20°C in less than 3 min and maintain target temperature to within ±0.3°C indefinitely. Here we describe cooling chip fabrication and performance in mediating cessation of neural signaling in acute preparations of rodents, ferrets, and primates. [ABSTRACT FROM AUTHOR]

Packaging continues to be one of the most challenging steps in micro-nanofabrication, as many emerging techniques (e.g., soft lithography) are incompatible with the standard high-precision alignment and bonding equipment. In this paper, we present a simple-to-operate, easy-to-adapt packaging strategy, referred to as Capillary-driven Automatic Packaging (CAP), to achieve automatic packaging process, including the desired features of spontaneous alignment and bonding, wide applicability to various materials, potential scalability, and direct incorporation in the layout. Specifically, self-alignment and self-engagement of the CAP process induced by the interfacial capillary interactions between a liquid capillary bridge and the top and bottom substrates have been experimentally characterized and theoretically analyzed with scalable implications. High-precision alignment (of less than 10 µm) and outstanding bonding performance (up to 300 kPa) has been reliably obtained. In addition, a 3D microfluidic network, aligned and bonded by the CAP technique, has been devised to demonstrate the applicability of this facile yet robust packaging technique for emerging microfluidic and bioengineering applications. [ABSTRACT FROM AUTHOR]

In this paper we describe a microfabrication-derived approach for defining interactions between distinct groups of cells and integrating biosensors with cellular micropatterns. In this approach, photoresist lithography was employed to micropattern cell-adhesive ligand (collagen I) on silane-modified glass substrates. Poly(ethylene glycol) (PEG) photolithography was then used to fabricate hydrogel microstructures in registration with existing collagen I domains. A glass substrate modified in this manner had three types of micrpatterned regions: cell-adhesive collagen I domains, moderately adhesive silanized glass regions, and nonadhesive PEG hydrogel regions. Incubation of this substrate with primary rat hepatocytes or HepG2 cells resulted in attachment of hepatic cells on collagen I domains with no adhesion observed on silane-modified glass regions or hydrogel domains. 3T3 fibroblasts added onto the same surface attached on the glass regions around the hepatocytes, completing the coculture. Significantly, PEG hydrogel microstructures remained free of cells and were used to “fence” hepatocytes from fibroblasts, thus limiting communication between the cell types. We also demonstrated that entrapment of enzyme molecules inside hydrogel microstructures did not compromise nonfouling properties of PEG. Building on this result, horse radish peroxidase-containing hydrogel microstructures were integrated into micropatterned cocultures and were used to detect hydrogen peroxide in the culture medium. The surface micropatterning approach described here may be used in the future to simultaneously define and detect endocrine signaling between two distinct cell types. [ABSTRACT FROM AUTHOR]

In this paper, a nanocomposite approach is introduced to provide both electrically conductive and optically transparent micropatterns on any flexible substrate employing photolithography-based microfabrication. The nanocomposite materials combine the highly directional nanoscopic networks and electrical conductivity of single-wall carbon nanotubes (SWNTs) with the photopatternability and optical transparency of SU-8 photoresist. The photopatternable nanocomposites have yielded high optical transparency of 90% and high electrical conductivity of 27.5 S m[?]1 with the minimal feature resolution of 10 um. Additionally, an interesting nano-bridge phenomenon has been discovered during fabrication of the microscale features. Moreover, the photopatternable transparent conductive nanocomposite has demonstrated its application to biomedical sensing for exceptional adaptability and flexibility. [ABSTRACT FROM AUTHOR]