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Biomedical subjects

Marc J Madou

Publications and source records attributed to Marc J Madou.

7 recordsLinked to original sources

3-D electrode designs for flow-through dielectrophoretic systems.

Traditional methods of dielectrophoretic separation using planar microelectrodes have a common problem: the dielectrophoretic force, which is proportional to nabla|E|2, rapidly decays as the distance from the electrodes increases. Recent advances in carbon microelectromechanical systems have allowed researchers to create carbon 3-D structures with relative ease. These developments have opened up new possibilities in the fabrication of complex 3-D shapes. In this paper, the use of 3-D electrode designs for high-throughput dielectrophoretic separation/concentration/filtration systems is investigated. 3-D electrode designs are beneficial because (i) they provide a method of extending the electric field within the fluid. (ii) The 3-D electrodes can be designed so that the velocity field coincides with the electric field distribution. (iii) Novel electrode designs, not based on planar electrodes designs, can be developed and used. The electric field distribution and velocity fields of 3-D electrode designs that are simple extensions of 2-D designs are presented, and two novel electrode designs that are not based on 2-D electrode designs are introduced. Finally, a proof-of-concept experimental device for extraction of nanofibrous carbon from canola oil is demonstrated.

Carbon↗

Genetically engineered protein in hydrogels tailors stimuli-responsive characteristics.

Certain proteins undergo a substantial conformational change in response to a given stimulus. This conformational change can manifest in different manners and result in an actuation, that is, catalytic or signalling event, movement, interaction with other proteins, and so on. In all cases, the sensing-actuation process of proteins is initiated by a recognition event that translates into a mechanical action. Thus, proteins are ideal components for designing new nanomaterials that are intelligent and can perform desired mechanical actions in response to target stimuli. A number of approaches have been undertaken to mimic nature's sensing-actuating process. We now report a new hybrid material that integrates genetically engineered proteins within hydrogels capable of producing a stimulus-responsive action mechanism. The mechanical effect is a result of an induced conformational change and binding affinities of the protein in response to a stimulus. The stimuli-responsive hydrogel exhibits three specific swelling stages in response to various ligands offering additional fine-tuned control over a conventional two-stage swelling hydrogel. The newly prepared material was used in the sensing, and subsequent gating and transport of biomolecules across a polymer network, demonstrating its potential application in microfluidics and miniaturized drug-delivery systems.

Calcium↗

Whole-cell-reporter-gene-based biosensing systems on a compact disk microfluidics platform.

Biosensing systems such as reporter-gene-based whole-cell assays are increasingly finding applications in biological and environmental screening. A whole-cell approach to such analyses can provide valuable information about the bioavailable level of a compound of interest. These biosensing systems rely on the molecular recognition of a specific analyte by a regulatory protein and, therefore, can detect low levels of the target analyte. In this study, Escherichia coli cells containing plasmid pSD10 were engineered to sense the model target analytes arsenite and antimonite, the target analytes in this study. The biosensing system takes advantage of the recognition of the regulatory protein, ArsR, for arsenite and antimonite to produce the reporter protein, which in this case is GFPuv. The fluorescence emitted by the GFPuv in the cells can be directly related to the concentration of the analyte in the cell, making this biosensing system useful in the detection of arsenite and/or antimonite in a variety of samples. Miniaturization of biosensing systems can further enhance their utility by decreasing reagent consumption and analysis time and by allowing for the high-throughput screening of samples. To that end, we employed a microcentrifugal microfluidics platform that has low power, space, and reagent requirements, increased speed of detection, and the potential for portability. Herein, we demonstrate for the first time the adaptation of a whole-cell sensing system to a microcentrifugal microfluidics platform. Moreover, we were able to detect our target analytes in a rapid and sensitive manner compared to conventional sensing methods.

Antimony↗

Cell lysis on a microfluidic CD (compact disc).

Cell lysis was demonstrated on a microfluidic CD (Compact Disc) platform. In this purely mechanical lysis method, spherical particles (beads) in a lysis chamber microfabricated in a CD, cause disruption of mammalian (CHO-K1), bacterial (Escherichia coli), and yeast (Saccharomyces cerevisiae) cells. Interactions between beads and cells are generated in the rimming flow established inside a partially filled annular chamber in the CD rotating around a horizontal axis. To maximize bead-cell interactions in the lysis chamber, the CD was spun forward and backwards around this axis, using high acceleration for 5 to 7 min. Investigation on inter-particle forces (friction and collision) identified the following parameters; bead density, angular velocity, acceleration rate, and solid volume fraction as having the most significant contribution to cell lysis. Cell disruption efficiency was verified either through direct microscopic viewing or measurement of the DNA concentration after cell lysing. Lysis efficiency relative to a conventional lysis protocol was approximately 65%. In the long term, this work is geared towards CD based sample-to-answer nucleic acid analysis which will include cell lysis, DNA purification, DNA amplification, and DNA hybridization detection.

Animals↗

Design of a compact disk-like microfluidic platform for enzyme-linked immunosorbent assay.

This paper presents an integrated microfluidic device on a compact disk (CD) that performs an enzyme-linked immunosorbent assay (ELISA) for rat IgG from a hybridoma cell culture. Centrifugal and capillary forces were used to control the flow sequence of different solutions involved in the ELISA process. The microfluidic device was fabricated on a plastic CD. Each step of the ELISA process was carried out automatically by controlling the rotation speed of the CD. The work on analysis of rat IgG from hybridoma culture showed that the microchip-based ELISA has the same detection range as the conventional method on the 96-well microtiter plate but has advantages such as less reagent consumption and shorter assay time over the conventional method.

Animals↗

Fluorescent ion-selective optode membranes incorporated onto a centrifugal microfluidics platform.

The development of an integrated analysis system for small ions based on ion-selective optodes and centrifugal microfluidics is reported. The performance of this system was evaluated through five-point calibration plots for two types of optode membranes, one being cation-selective and the other anion-selective, which were incorporated into a microfluidics platform on which fluid motion is induced via angular rotation. Additionally, the application of the microfluidic platform to ion analysis is studied via a two-point calibration protocol used to quantify an unknown sample. Calibrant solutions are delivered from reservoirs fabricated onto the platform to a measuring area that contains the optode membrane, with a change in membrane fluorescence being monitored. This work demonstrates the first instance of a microfluidic-based analysis system with detection based on ion-selective optode membranes monitored with fluorescence transduction. Furthermore, in addition to employing a standard excitation source where a fiber-optic probe is coupled to a tungsten-halogen lamp, laser diodes such as those employed in portable CD/DVD players were studied as excitation sources to enhance the observed fluorescence signals.

Journal Article↗