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Biomedical subjects

David J Beebe

Publications and source records attributed to David J Beebe.

At least 19 recordsLinked to original sources

Magnetically-driven biomimetic micro pumping using vortices.

Planar micropumps utilizing vortices shed by an oscillating ferromagnetic bar are presented. The movement of the bar is induced by magnetic coupling with an external spinning magnet. Thus, energy transfer is achieved without physical contact or need of any on-chip power source. To create vortices inside the chip, the Reynolds number is locally increased with the oscillation of the bar. The utilization of the vortices as a tool for efficient transfer of energy is inspired by the behaviour of swimming animals and flying insects in nature. The pumps operate in two different scales (milli-scale and micro-scale) in order to take advantage of both. Experiments are presented characterizing the pumps and their flow patterns. The range of operation of the pumps is from 3 microl min(-1) to 600 microl min(-1), though smaller flow rates are also possible.

Biomimetics↗

Hydrodynamic microfabrication via"on the fly" photopolymerization of microscale fibers and tubes.

A microfluidic apparatus capable of creating continuous microscale cylindrical polymeric structures has been developed. This system is able to produce microstructures (e.g. fibers, tubes) by employing 3D multiple stream laminar flow and "on the fly"in-situ photopolymerization. The details of the fabrication process and the characterization of the produced microfibers are described. The apparatus is constructed by merging pulled glass pipettes with PDMS molding technology and used to manufacture the fibers and tubes. By controlling the sample and sheath volume flow rates, the dimensions of the microstructures produced can be altered without re-tooling. The fiber properties including elasticity, stimuli responsiveness, and biosensing are characterized. Responsive woven fabric and biosensing fibers are demonstrated. The fabrication process is simple, cost effective and flexible in materials, geometries, and scales.

Acrylates↗

An externally driven magnetic microstirrer.

In this paper, an inexpensive, easy-to-fabricate active magnetic mixer is presented. This mixer functions on top of a common magnetic stir plate and is capable of mixing two streams, each at flow rates up to 5 ml min(-1). A liquid-phase photopolymerization technique is used to fabricate the device. An analysis of mixing efficiency is based on greyscale intensity measurements of two coloured streams passing through the mixer. A brief hypothesis of the mechanism of mixing is also presented.

Complex Mixtures↗

Embryonic development in the mouse is enhanced via microchannel culture.

Microfluidic devices (microchannels) have been fabricated and tested for embryo culture. Three different microfabrication materials (silicon, polydimethylsiloxane (PDMS), and borosilicate) were used to fabricate the microchannels. The objective of this study was to determine if static microchannels permitted culture of mouse embryos to the blastocyst stage. Groups of 10 two-cell ICR x B6SJL/F1 mouse embryos were cultured for 96 hours in 4 different physical culture systems: 1) silicon/borosilicate microchannels, 2) PDMS/borosilicate microchannels, and 3) standard microdrops. Embryos cultured in the silicon/borosilicate and PDMS/borosilicate microchannels exhibited a faster rate of cleavage (P < 0.05), and produced more blastocysts (P < 0.01) than control microdrops. Furthermore, microchannels had a lower percentage of degenerated embryos than control embryos (P < 0.01). The results suggest that the microchannel culture systems may provide a culture environment that more closely mimics the in vivo environment.

Animals↗

Flow control with hydrogels.

With the advent of the genomic revolution and the sequencing of the human genome complete, the majority of pharmaceuticals under development are proteins. Consequently, new techniques to more effectively administer these new protein therapeutics need to be developed. One method that is gaining popularity in the research community involves the use of responsive hydrogel actuators for flow control in drug delivery devices. Responsive hydrogels are materials able to undergo a volume change in response to a stimulus from their local environment. The following paper overviews recent advances made using hydrogel actuators for flow control such as resistance based valves, hydrogel jacket valves, hybrid hydrogel membrane valve, electrically triggered valves, and biomimetic valves. Also reviewed are several hydrogel flow control systems such as a flow sorter and pH-regulation system. The chemistry of the hydrogel actuators can be tweaked to allow physiological variables to trigger the volume expansion of the hydrogel actuators as demonstrated by several glucose sensitive hydrogel valves reviewed below. Therefore, the door to physiological feedback controlling the infusion rate in a drug delivery device is opened and has the potential to revolutionize protein pharmaceutical drug delivery.

Drug Delivery Systems↗

Microenvironment design considerations for cellular scale studies.

In vivo cellular microenvironments are not well-mimicked in present in vitro cell culture systems. Microtechnology, and microfluidics in particular, provides the tools to create in vivo-like cellular microenvironments in vitro. Features of in vitro cellular microenvironments are discussed and compared to macroscale cell culture environments; the concept of an effective culture volume (ECV) is introduced to facilitate the comparison. Current research using microtechnology to investigate in vitro cellular microenvironments is presented and areas where more research is needed in characterizing the in vitro microenvironment are outlined.

Adsorption↗

Microfluidic tectonics platform: A colorimetric, disposable botulinum toxin enzyme-linked immunosorbent assay system.

A fabrication platform for realizing integrated microfluidic devices is discussed. The platform allows for creating specific microsystems for multistep assays in an ad hoc manner as the components that perform the assay steps can be created at any location inside the device via in situ fabrication. The platform was utilized to create a prototype microsystem for detecting botulinum neurotoxin directly from whole blood. Process steps such as sample preparation by filtration, mixing and incubation with reagents was carried out on the device. Various microfluidic components such as channel network, valves and porous filter were fabricated from prepolymer mixture consisting of monomer, cross-linker and a photoinitiator. For detection of the toxoid, biotinylated antibodies were immobilized on streptavidin-functionalized agarose gel beads. The gel beads were introduced into the device and were used as readouts. Enzymatic reaction between alkaline phosphatase (on secondary antibody) and substrate produced an insoluble, colored precipitate that coated the beads thus making the readout visible to the naked eye. Clinically relevant amounts of the toxin can be detected from whole blood using the portable enzyme-linked immunosorbent assay (ELISA) system. Multiple layers can be realized for effective space utilization and creating a three-dimensional (3-D) chaotic mixer. In addition, external materials such as membranes can be incorporated into the device as components. Individual components that were necessary to perform these steps were characterized, and their mutual compatibility is also discussed.

Antibodies↗

Developments in in vitro technologies for swine embryo production.

Several modifications have been made to in vitro production (IVP) systems to allow more efficient production of viable porcine embryos. Although in vitro production of pig embryos has been studied for over 30 years, the overall blastocyst production rate remains low. The low blastocyst rate is due to several factors, including polyspermic oocyte penetration, low rate of male pronucleus formation and less than optimal in vitro culture systems. These conditions are all inherent problems in porcine IVP and many of the mechanisms involved remain unknown. Considerable research has examined culture medium and the techniques used during the various stages of in vitro production. However, changes to the physical culture system used during IVF have remained unchanged until recently. The present paper will summarise selected developments in fertilisation and embryo culture media composition and focus on the development of modified equipment to improve the conditions used during the IVP of porcine oocytes and embryos.

Animals↗

Microfluidic valve with cored glass microneedle for microinjection.

In this paper, a new microinjection device was constructed by fusing a glass microneedle and a PDMS-based microvalve. The microneedle was fabricated via traditional micropipette pulling. The PDMS-based microvalve regulates the fluid flow in the microchannel and microneedle. The 'ON/OFF' operation of the valve was controlled by manually supplied pneumatic pressure. The valve membrane utilized a two level geometry to improve control at low flow rates. The relation between pressure and flow was measured and the results showed that very small volumes of fluid (>1 nl) could be controlled. The valve operation was investigated by monitoring the tip of the needle and pneumatic pressure simultaneously and it demonstrated very stable 'ON/OFF' operation to the pressure change.

Dimethylpolysiloxanes↗

In situ fabricated porous filters for microsystems.

Porous filters were fabricated inside a microchannel using emulsion photo-polymerization. The porosity of the filter is dependent on the composition of the pre-polymer mixture. The morphology and flow resistance of four filters with varying water and cross-linker concentrations was studied. The porous filter was used to separate samples, such as whole blood to cell/serum components, based on size. The efficiency of cell/serum separation by the porous filter was tested by performing a glucose-6-phosphate dehydrogenase assay and was found to be comparable to separation by centrifuge. The porous filter thus mimics the functionality of the centrifuge, with the added benefits of no power requirements and ability to handle small sample volumes.

Acetophenones↗

Principles of surface-directed liquid flow in microfluidic channels.

To direct liquid flow inside microchannels, surface free energies were patterned by use of self-assembled monolayers (SAMs) in combination with either multistream laminar flow or photolithography. For the photolithographic method, two photocleavable SAMs were designed and synthesized. Carboxylic acid-terminated monolayers were obtained by photodeprotection, which was confirmed by contact angle and X-ray photoelectron spectroscopy. Using either of these patterning methods, we show that aqueous liquids flow only along the hydrophilic pathways when the pressure is maintained below a critical value; the liquids are referred to as being confined by virtual walls. Several principles of liquid flow in surface-patterned channels were derived analytically and verified experimentally. These principles include the maximum pressure that virtual walls can withstand, the critical width of the hydrophilic pathway that can support spontaneous flow, the smallest width of the liquid streams under an external pressure, the critical radius of curvature of turns that can be introduced into the hydrophilic pathway without liquid crossing the hydrophilic-hydrophobic boundary, and the minimal distance for two liquid streams to remain separated under the maximum pressure. Experimental results are in good agreement with the analytical predictions.

Biotechnology↗

A passive pumping method for microfluidic devices.

The surface energy present in a small drop of liquid is used to pump the liquid through a microchannel. The flow rate is determined by the volume of the drop present on the pumping port of the microchannel. A flow rate of 1.25 microL s(-1) is demonstrated using 0.5 microL drops of water. Two other fluid manipulations are demonstrated using the passive pumping method: pumping liquid to a higher gravitational potential energy and creating a plug within a microchannel.

Journal Article↗

Control and applications of immiscible liquids in microchannels.

Photolithography was used in combination with photocleavable self-assembled monolayers to pattern surface free energies inside microchannels enabling the control of the boundary between immiscible liquids. While aqueous solutions are confined to the hydrophilic pathways by surface forces alone, organic liquids are confined to the hydrophobic region only if the aqueous liquid first occupies the hydrophilic region. In this way, stable liquid boundaries between immiscible liquids are possible as long as the pressures are maintained below critical values. The maximum pressures are determined by the interfacial tension of the aqueous solution and organic liquid, channel depth, and advancing contact angle (theta;(a)). Experimental results on maximum pressures are in good agreement with the analytical values. The ability to confine and position the boundary between immiscible liquids inside microchannels leads to a broad range of applications in microfluidic systems, which is exemplified by fabrication of a semipermeable membrane in a surface-patterned channel via interfacial polymerization.

Journal Article↗

An evaporation-based microfluidic sample concentration method.

We present a method for sample concentration within microfluidic devices using evaporation-induced flow. Evaporation-induced flow is easy to incorporate into microfluidic designs and can be used to concentrate a wide variety of molecules. The practicality of this method was demonstrated with 0.2 microm fluorescent spheres and FITC-labeled BSA. Thirty two percent of the 0.6 microL fluorescent sphere suspension was concentrated into a well within a microfluidic device. In the same amount of time, 93% of the 0.6 microL FITC-labeled BSA solution was concentrated.

Journal Article↗

Physics and applications of microfluidics in biology.

Fluid flow at the microscale exhibits unique phenomena that can be leveraged to fabricate devices and components capable of performing functions useful for biological studies. The physics of importance to microfluidics are reviewed. Common methods of fabricating microfluidic devices and systems are described. Components, including valves, mixers, and pumps, capable of controlling fluid flow by utilizing the physics of the microscale are presented. Techniques for sensing flow characteristics are described and examples of devices and systems that perform bioanalysis are presented. The focus of this review is microscale phenomena and the use of the physics of the scale to create devices and systems that provide functionality useful to the life sciences.

Biosensing Techniques↗