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Microfluidic arrays in genetic analysis.

The goal of genetic analysis is to discover genetic markers that are informative for providing high confidence, positive predictive value in managing phenotypic outcomes. Primary consensus sequence data, genetic polymorphism databases and associated phenotype data are rapidly making genetic analysis more useful. Therefore, genetic analysis applications are gradually becoming more mainstream. The diversity and complexity of genetic analysis currently requires an array of analytical techniques, instrument platforms and software to support all the steps from data acquisition to interpretation. As supporting research technologies mature, they are incorporating increasing levels of automation, system integration and miniaturization. Microfluidic arrays are positioned to play a key role in routine genetic analysis, particularly as they begin to appear in more fully integrated analytical platforms.

Animals↗

Integrated microfluidics for chromosome engineering--preparation, transportation and manipulation.

The design and fabrication process of microfluidics for chromosome transportation and manipulation are described. Micro-channels for fluid networks could be built on single-crystal silicon substrates by anisotropic wet etching. After injection of a chromosome-containing buffer solution into the channels by a mechanical pump, chromosome electrophoresis was carried out for its precise transportation. The behavior of chromosomes placed in direct (DC) and alternating (AC) electric fields is explained in detail. The frequency and size dependent amplitude of the chromosome observed in this study showed the existence of inertial hydrodynamic effects in an oscillatory motion. Using a rotating AC electric field, chromosomes could be rotated due to the induced polarization of surface charge and electric double layer of the chromosomes. This study thus shows the possibility of chromosome engineering on microchips, which is expected to enable the development of new devices for chromosome research.

Biophysical Phenomena↗

Quantitative detection and fixation of single and multiple gold nanoparticles on a microfluidic chip by thermal lens microscope.

A detection and fixation method of single and multiple gold nanoparticles on the wall of a microfluidic channel is demonstrated. A thermal lens microscope (TLM) with continuous-wave excitation (wavelength, 532 nm) and probe (wavelength, 670 nm) laser beams was used to realize the sensitive detection of heat generated by light absorption of individual gold nanoparticles (50 nm in diameter); fixation of the individual nanoparticles was realized simultaneously. The fixation mechanism was investigated and attributed to an absorption-based optical force. In addition to single nanoparticle detection, multiple-nanoparticle detection and fixation was demonstrated. An acceleration of fixation was observed when the number of fixed particles was increased. TLM is expected to be a powerful tool for both the quantitative detection and precise fixation of individual nanoparticles.

Journal Article↗

Performance of in-line microfluidic mixers in laminar flow for high-throughput flow cytometry.

We describe a micromixing approach that is compatible with commercial autosamplers, flow cytometry, and other detection schemes that require the mixing of components that have been introduced into laminarflow. The scheme is based on high-throughput flow cytometry (HyperCyt) where samples from multi-well plates that have been picked up by an autosampler can be separated during delivery by the small air bubbles introduced during the transit of the autosampler probe from well to well. Here, either cell or particle samplesflowing continuously and driven by a syringe are brought together in a Y with reagent samples from wells driven by a peristaltic pump. The mixing is driven by a magnetic microstirrer contained within the sample line. The mixing is assessed using fluorescence of both cell calcium responses and bead-based fluorescence unquenching. In the analysis stream, the particles and reagents are mixed with eithera "wire" or "bar". The bar is more efficient than the wire, and the efficiency of either depends on the spinning action. The high-throughput approach and mixing in HyperCyt integrate autosamplers with submicroliter detection volumes for analysis in flow cytometry or in microfluidic channels.

Equipment Design↗

Large-scale blood substitute production using a microfluidizer.

Microfluidization has been tested as a way to disperse phospholipids in aqueous hemoglobin solutions. Spherical and stable liposomes of 2 to 3 microns were obtained. Lipid incorporation (up to 85%) and hemoglobin encapsulation (up to 15%) in liposomes have been improved with respect to previous investigations. However, results show that a more efficient dispersion system using lower concentrations of lipid is required to obtain a high liposome hemoglobin concentration (limited actually to 150 g/l) and an economically and biologically suitable process for artificial blood production at large scale.

Blood Substitutes↗

[Lab-on-a-chip (microfluidics) technology].

Over the last decade, the lab-on-a-chip concept has received considerable attention. This technology promises significant advantages in terms of speed, cost, sample/reagent consumption, contamination, efficiency and automation. In this review, we focus on lab-on-a-chip with fluid flow along micro-scale channels (microfluidics) and its application on life science and biotechnology.

Chemistry Techniques, Analytical↗

Microfluidic systems for high-throughput and combinatorial chemistry.

Demands on modern combinatorial chemistry have necessitated massive investment in new synthetic technologies that will allow the production of highly pure drug candidates on short timescales. The advent of microfluidic reactor technologies has recently provided a new tool for the combinatorial chemist, offering the promise of ultra-high-throughput solution-phase chemistries. This review describes why such microscale systems provide unique environments for performing high-efficiency molecular synthesis, details some of the most important recent developments in the field and assesses the true applicability of micro-engineered reactors in high-throughput compound synthesis.

Combinatorial Chemistry Techniques↗

Disposable microfluidic ELISA for the rapid determination of folic acid content in food products.

A micro-analytical system for rapid and quantitative analysis by inhibition immunoassay is presented and applied to the detection of folic acid. Eight polymer microchannels of 65-nL volume each and containing microelectrodes are embedded in a cartridge so that they can be operated simultaneously. All fluidic steps as well as the amperometric detection in the channels are operated by an instrument and software developed in-house. The fluidic steps of the immunoassay occur through hydrodynamic loading of the different solutions through the channels. The speed and duration of the flow and incubation parameters can thus be adapted to the biological and testing requirements. The effectiveness of the system was demonstrated by analysing folic acid concentrations in real infant formula samples within 5 min. In an effort to get a fully monitored assay, each fluidic step is monitored thanks to continuous amperometric detection of oxygen in the microchannel.

Alkaline Phosphatase↗

Volume cytometry: microfluidic sensor for high-throughput screening in real time.

Regulation of cell volume was one of the earliest evolutionary demands for life and remains a universal measure of cell metabolism. Since conventional methods to measure cell volume, such as microscopy, are complex and time-consuming, cell volume has not been used as the basis for cell-based screening. We have developed a microfabricated chip that can measure the volume of small numbers of cells in real time with unprecedented resolution. The method is applicable to adherent or suspended populations of cells and membrane-bound organelles. Our prototype device can detect volume changes in a monolayer of tissue-cultured astrocytes responding to anisotonic stimuli of <1mOsm. We determined the sensitivity to antibiotics of different E. coli strains in <10 min at 24 degrees C. This time can be reduced at higher temperatures enabling on-site clinical testing of infectious agents. Using the chip to screen natural products, we found a peptide in spider venom that inhibits eukaryotic volume regulation at approximately 100pM. The prototype chip made in silicon is inexpensive, reusable, and runs on low-voltage electrical power. The technology can be readily transferred to large arrays in plastic.

Algorithms↗

Analysis of non-Newtonian liquids using a microfluidic capillary viscometer.

We have used a self-calibrating microfabricated capillary viscometer to analyze non-Newtonian power law fluids. The capillary viscometer can generate a wide range of shear rates during a single experimental run enabling quick and accurate analysis of non-Newtonian liquids. The measurement of viscosity is based on monitoring the capillary pressure-driven movement of fluid sample whose mean velocity and, therefore, shear rate varies with time. The device has been successfully tested for accuracy and robustness with dilute as well as semidilute solutions of flexible elastic polymers including poly(ethylene oxide) and hydrolyzed polyacrylamide to an aqueous solution of a stiff rodlike polymer molecule of xanthan gum, a popular emulsifier and food thickener, as well as with ink-jet printing inks. Viscosities in the range of 1-600 cP were measured, and shear rates varying from 5 to 1000 s(-1) have been obtained on the microfabricated viscometer with the current geometry and channel dimensions. The total measurement time varied between 2 and 8 min and less than 1 microL of sample volume was required. Such a microfabricated capillary viscometer would have possible applications in quality control and manufacturing where rapid and repeated measurements need to be made using limited sample volume.

Biopolymers↗

Micro magnetic stir-bar mixer integrated with parylene microfluidic channels.

Previously, we reported a micro magnetic stir-bar mixer driven by an external rotating magnetic field and its rapid mixing performance in polydimethyl-siloxane (PDMS) channels. The PDMS piece with embedded fluid channels were manually aligned to a glass substrate and assembled. In this paper, we report the fabrication and testing results of a micro magnetic stir-bar monolithically integrated in parylene surface-micromachined channels with improved design features, including small tolerance of the stir-bar to channel wall (10 microm). Using of parylene based microchannels with improved design not only provides improved mixing, but also eliminates certain problems associated with PDMS-based channels. For example, porosity of PDMS causes evaporation and absorption of chemicals and thus channels made of PDMS are prone to cross-contamination. We have also demonstrated that the magnetic stir-bar can be used to pump liquid in micro channels.

Dimethylpolysiloxanes↗

Reduction of polyspermic penetration using biomimetic microfluidic technology during in vitro fertilization.

Efforts to improve the in vitro embryo production process in pigs have included modifying culture medium and number of spermatozoa inseminated in order to reduce the incidence of polyspermy. Polyspermy is a pathological condition which results in aberrant embryonic development. The microchannels are designed to more closely mimic the function of the oviduct and create a flow pattern of spermatozoa past the oocytes similar to the pattern in the oviduct. In vitro fertilization of porcine oocytes in the microchannels has produced a higher incidence of monospermic penetration (p<0.05) as compared to the oocytes fertilized in the traditional microdrop system with comparable penetration and male pronucleus formation rates. Additionally, cleavage rates of the embryos as well as development to the blastocyst stage are similar. Here we demonstrate that the biomimetic microchannel in vitro fertilization system can reduce polyspermy and, therefore, increase the number of potentially viable embryos without reducing the overall in vitro production efficiency.

Animals↗