Lawrence Kulinsky

Nanopore Technology for Biomedical Applications

The ability to create well-defined and controlled interfaces has been an area of great interest over the last few years, particularly in the biomedical arena. This paper will describe the development of technology for the fabrication of nanopore membranes, and their operation in biological environments. With monodisperse pores sizes as small as 10 nanometers, these membranes offer advantages in their reproducibility, and their ability to be integrated with controlled biochemical surface modification protocols. A comprehensive review of results in the areas of nanopore and biocapsule microfabrication technologies, biocompatibility of nanomembrane materials, biologically appropriate post-processing protocols (bonding, sterilization), surface modification protocols, and appropriate mass transport models will be presented. The results point to the potential of using such technologies for therapeutic applications including immunoisolation biocapsules, drug delivery devices, and targeted biorecognition platforms.

Controlled Continuous Patterning of Polymeric Nanofibers on Three-Dimensional Substrates Using Low-Voltage Near-Field Electrospinning

We report on a continuous method for controlled electrospinning of polymeric nanofibers on two-dimensional (2D) and three dimensional (3D) substrates using low voltage near-field electrospinning (LV NFES). The method overcomes some of the drawbacks in more conventional near-field electrospinning by using a superelastic polymer ink formulation. The viscoelastic nature of our polymer ink enables continuous electrospinning at a very low voltage of 200 V, almost an order of magnitude lower than conventional NFES, thereby reducing bending instabilities and increasing control of the resulting polymer jet. In one application, polymeric nanofibers are freely suspended between microstructures of 3D carbon on Si substrates to illustrate wiring together 3D components in any desired pattern.

Carbon post-microarrays for glucose sensors

A novel design and fabrication method of glucose sensors based on high aspect ratio carbon post-microarrays is reported in this paper. Apart from the fact that carbon has a wide electrochemical stability window, a major advantage of using carbon post-microarrays as working electrodes for an amperometric glucose sensor is the large reactive surface per unit footprint substrate area, improving sensitivity of the glucose sensor. The carbon post-microarrays were fabricated by carbon-microelectromechanical systems (C-MEMS) technology. Immobilization of enzyme onto the carbon post-electrodes was carried out through co-deposition of glucose oxidase (GOx) and electrochemically polymerized polypyrrole (PPy). Sensing performance of the glucose sensors with different post-heights and various post-densities was tested and compared. The carbon post-glucose sensors show a linear range from 0.5mM to 20mM and a response time of about 20s, which are comparable to the simulation result. Sensitivity per unit footprint substrate area as large as 2.02mA/(mMcm(2)) is achieved with the 140microm high (aspect ratio around 5:1) carbon post-samples, which is two times the sensitivity per unit footprint substrate area of the flat carbon films. This result is consistent with the hypothesis that the number of reaction sites scales with the reactive surface area of the sensor. Numerical simulation based on enzymatic reaction and glucose diffusion kinetics gives the optimum geometric design rules for the carbon post-glucose sensor. Glucose sensors with even higher sensitivity can be achieved utilizing higher carbon post-microarrays when technology evolution will permit it.

Theoretical development and critical analysis of burst frequency equations for passive valves on centrifugal microfluidic platforms.

This paper presents a theoretical development and critical analysis of the burst frequency equations for capillary valves on a microfluidic compact disc (CD) platform. This analysis includes background on passive capillary valves and the governing models/equations that have been developed to date. The implicit assumptions and limitations of these models are discussed. The fluid meniscus dynamics before bursting is broken up into a multi-stage model and a more accurate version of the burst frequency equation for the capillary valves is proposed. The modified equations are used to evaluate the effects of various CD design parameters such as the hydraulic diameter, the height to width aspect ratio, and the opening wedge angle of the channel on the burst pressure.

Lab-on-a-CD A Fully Integrated Molecular Diagnostic System

The field of centrifugal microfluidics has experienced tremendous growth during the past 15 years, especially in applications such as lab-on-a-disc (LoD) diagnostics. The strength of LoD systems lies in its potential for development into fully integrated sample-to-answer analysis systems. This review highlights the technologies necessary to develop the next generation of these systems. In addition to outlining valving and other fluid-handling operations, we discuss the recent advances and future outlook in four categories of LoD processes: reagent storage, sample preparation, nucleic acid amplification, and analyte detection strategies.

3-D Micro and Nano Technologies for Improvements in Electrochemical Power Devices

This review focuses on recent advances in micro- and nano-fabrication techniques and their applications to electrochemical power devices, specifically microfabricated Lithium-ion batteries, enzymatic and microbial fuel cells (biofuel cells), and dye-sensitized solar cells (DSSCs). Although the maturity of these three technologies ranges from market ready (batteries) to fundamental research (biofuel cells) to applied research (DSSCs), advances in MEMS (Micro-Electro-Mechanical Systems) and NEMS (Nano-Electro-Mechanical Systems) techniques, particularly modifications in surface area and surface chemistry, and novel genetic and molecular engineering techniques, significantly improve the electrochemical activity of these technologies across the board. For each of these three categories of power-MEMS devices the review covers: (1) The technical challenges facing the performance and fabrication of electrochemical power devices; (2) Current MEMS and NEMS techniques used to improve efficiency; and (3) Future outlook and suggested improvements of MEMS and NEMS for implementation in electrochemical power devices.

Electroviscous effects in microchannels

Fluid flow in capillary microchannels is used in numerous applications in biotechnology (such as protein separation, fast DNA analysis, drug deliveries systems and viral filtration), in solid-state devices, and in catalytic devices. The current work presents the experimental validation for the electrokinetic theory in microchannels. Retardation of polar liquids, including de-ionized water, ethanol and propyl alcohol, is studied in microfabricated channels of several diameters. It was found that polar liquids flow about 6 percent more slowly than predicted by the classical hydrodynamic theory in microchannels, with the hydraulic diameter equal to 90 microns. For small microchannels with a hydraulic diameter of several microns, observed retardation is on the order of 70 percent. Collected experimental data have good correspondence with the electrokinetic model presented. Electrokinetic retardation of polar liquids in microchannels is based on the charge separation principle. Electrical charges are separated at the interface (near the channel wall). When liquid is forced downstream, it causes charge accumulation at one end of the microchannel. The streaming potential produced causes an upstream current that creates upstream counterflow. The resultant fluid flow is less than it would be for non-polar liquids. The higher the zeta-potential at the microchannel wall and the smaller the channel, the larger the resulting retardation. Modifications for the friction factor, as applied to microfluidics, are suggested. Recommendations to improve fluid flow in microchannels are made.

Capillary filling in centrifugally actuated microfluidic devices with dynamically evolving contact line motion

In the present work, we analyze the capillary filling dynamics in centrifugally actuated microfluidic platforms with dynamically evolving contact line motion for wetting fluids. Special attention is devoted to estimate the effects of variable hydraulic resistances over different flow regimes. Dynamics of the meniscus advancement within the rotating microchannel turns out to be typically nonlinear, in tune with the relative instantaneous strengths of the capillary forces, centrifugal forces, and viscous resistances. Detailed dynamical characteristics of the meniscus evolution are obtained from the approximate semianalytical and full-scale numerical solutions, and are found to agree well with the experimental findings on lab-on-a-compact disk arrangements.

Diffusion-free mediator based miniature biofuel cell anode fabricated on a carbon-MEMS electrode.

We report on the functionalization of a micropatterned carbon electrode fabricated using the carbon-MEMS process for its use as a miniature diffusion-free glucose oxidase anode. Carbon-MEMS based electrodes offer precise manufacturing control on both the micro- and nanoscale and possess higher electron conductivity than redox hydrogels. However, the process involves pyrolysis in a reducing environment that renders the electrode surface less reactive and introduction of a high density of functional groups becomes challenging. Our functionalization strategy involves the electrochemical oxidation of amine linkers onto the electrode. This strategy works well with both aliphatic and aryl linkers and uses stable compounds. The anode is designed to operate through mediated electron transfer between 2,5-dihydroxybenzaldehyde (DHB) based redox mediator and glucose oxidase enzyme. The electrode was first functionalized with ethylene diamine (EDA) to serve as a linker for the redox mediator. The redox mediator was then grafted through reductive amination, and attachment was confirmed through cyclic voltammetry. The enzyme immobilization was carried out through either adsorption or attachment, and their efficiency was compared. For enzyme attachment, the DHB attached electrode was functionalized again through electro-oxidation of aminobenzoic acid (ABA) linker. The ABA functionalization resulted in reduction of the DHB redox current, perhaps due to increased steric hindrance on the electrode surface, but the mediator function was preserved. Enzyme attachment was then carried out through a coupling reaction between the free carboxyl group on the ABA linker and the amine side chains on the enzyme. The enzyme incubation for both adsorption and attachment was done either through a dry spotting method or wet spotting method. The dry spotting method calls for the evaporation of enzyme droplet to form a thin film before sealing the electrode environment, to increase the effective concentration of the enzyme on the electrode surface during incubation. The electrodes were finally protected with a gelatin based hydrogel film. The anode half-cell was tested using cyclic voltammetry in deoxygenated phosphate buffer saline solution pH 7.4 to minimize oxygen interference and to simulate the pH environment of the body. The electrodes that yielded the highest anodic current were prepared by enzyme attachment method with dry spotting incubation. A polarization response was generated for this anodic half-cell and exhibits operation close to maximum efficiency that is limited by the mass transport of glucose to the electrode.

Development of integrated protection for a miniaturized drug delivery system

A bi-layer structure comprising a thin gold layer and a polypyrrole (PPy) film is developed as a valve. To create such structures, the thin metal layer is used as a working electrode, and the polypyrrole film is electrochemically deposited on the metal electrode. A layer of Cr deposited under a stationary part of the bi-layer flap adheres strongly to the gold layer and anchors the hinge of the flap, while the layer of polyimide deposited under a movable part of the flap adheres weakly to the gold layer of the flap. This bi-layer flap structure functions as an actuator valve for the opening and closing of an aperture. The actuation mechanism of the valve is based on a volume change in the PPy layer as cations move in and out of the polymer film during reduction and oxidation reactions. A bias of 1.2 V is used to actuate the flap. A method is proposed to increase the drug release systems functionality by placing the drug release flap within a protective enclosure that also serves as a drug reservoir. This integrated protection ensures reliable operation of the drug release flap unencumbered by surrounding tissues when used in vivo. A prototype system using a PDMS drug reservoir has been successfully tested in PBS buffer solution. The proposed integrated protection system holds promise for implantable biomedical devices.