Chemical technology: all together now.
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Aspergillus↗
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Results for “Microchip Analytical Procedures”
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Lab on a chip: a little goes a long way.
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Chemistry↗
Soft X-ray microscopy at a spatial resolution better than 15 nm.
Analytical tools that have spatial resolution at the nanometre scale are indispensable for the life and physical sciences. It is desirable that these tools also permit elemental and chemical identification on a scale of 10 nm or less, with large penetration depths. A variety of techniques in X-ray imaging are currently being developed that may provide these combined capabilities. Here we report the achievement of sub-15-nm spatial resolution with a soft X-ray microscope--and a clear path to below 10 nm--using an overlay technique for zone plate fabrication. The microscope covers a spectral range from a photon energy of 250 eV (approximately 5 nm wavelength) to 1.8 keV (approximately 0.7 nm), so that primary K and L atomic resonances of elements such as C, N, O, Al, Ti, Fe, Co and Ni can be probed. This X-ray microscopy technique is therefore suitable for a wide range of studies: biological imaging in the water window; studies of wet environmental samples; studies of magnetic nanostructures with both elemental and spin-orbit sensitivity; studies that require viewing through thin windows, coatings or substrates (such as buried electronic devices in a silicon chip); and three-dimensional imaging of cryogenically fixed biological cells.
Biological Science Disciplines↗
Scaling and the design of miniaturized chemical-analysis systems.
Micrometre-scale analytical devices are more attractive than their macroscale counterparts for various reasons. For example, they use smaller volumes of reagents and are therefore cheaper, quicker and less hazardous to use, and more environmentally appealing. Scaling laws compare the relative performance of a system as the dimensions of the system change, and can predict the operational success of miniaturized chemical separation, reaction and detection devices before they are fabricated. Some devices designed using basic principles of scaling are now commercially available, and opportunities for miniaturizing new and challenging analytical systems continue to arise.
Animals↗
Future lab-on-a-chip technologies for interrogating individual molecules.
Advances in technology have allowed chemical sampling with high spatial resolution and the manipulation and measurement of individual molecules. Adaptation of these approaches to lab-on-a-chip formats is providing a new class of research tools for the investigation of biochemistry and life processes.
Lab-On-A-Chip Devices↗
Cells on chips.
Microsystems create new opportunities for the spatial and temporal control of cell growth and stimuli by combining surfaces that mimic complex biochemistries and geometries of the extracellular matrix with microfluidic channels that regulate transport of fluids and soluble factors. Further integration with bioanalytic microsystems results in multifunctional platforms for basic biological insights into cells and tissues, as well as for cell-based sensors with biochemical, biomedical and environmental functions. Highly integrated microdevices show great promise for basic biomedical and pharmaceutical research, and robust and portable point-of-care devices could be used in clinical settings, in both the developed and the developing world.
Animals↗
Single-cell growth and division dynamics showing epigenetic correlations.
The emergence of variation and subsequent inheritance of the emergent characteristics in a clonal population of bacteria is considered as evidence for epigenetic processes in the cell. We report here the results of experiments in which we quantitatively examined variations in single Escherichia coli cells with an identical genetic endowment in order to establish whether certain characteristics of single cells were inherited by their descendants maintained in a uniform environment. Significantly large variations of interdivision time, initial length, and final length were observed from generation to generation. Comparing the generations shows that interdivision time had no correlation with that of the consecutive generations, whereas those of initial length and final length were positively correlated with those of neighbouring generations.
Bacteriological Techniques↗
Amperometric determination of nitric oxide derived from pulmonary artery endothelial cells immobilized in a microchip channel.
A simple method for immobilizing a confluent layer of bovine pulmonary artery endothelial cells (bPAECs) in microchip-based channels is described. The microchips are prepared from poly(dimethylsiloxane) and have channel dimensions that approximate resistance vessels in vivo. The reversibly sealed channels were coated with fibronectin (100 microg ml(-1)) by aspiration. The bPAECs, which were introduced in the same manner, became attached to the fibronectin coating in about 2 h. The microchip could then be resealed over a micromolded carbon ink electrode (24 microm width x 6 microm height). Coating the carbon microelectrode with a 0.05% Nafion solution selectively blocked nitrite (10 microM) from being transported to the electrode surface while nitric oxide (NO, 10 microM) was amperometrically measured. Upon stimulation with adenosine triphosphate (ATP, 100 microM) the immobilized bPAECs produced and released micromolar amounts of NO. This NO production was effectively inhibited when the immobilized cells were incubated with L-nitro-arginine methyl ester (L-NAME), a competitive inhibitor for nitric oxide synthase. Moreover, once the immobilized bPAECs were no longer able to produce NO, incubation with L-arginine allowed for further ATP-stimulated NO production.
Animals↗
Separation of denatured proteins in free solution on a microchip based on differential binding of alkyl sulfates with different carbon chain lengths.
Denatured proteins were separated in free solution on a microfluidic chip without any sieving matrix after carrying out the denaturation with a mixture of alkyl sulfates with different carbon chain lengths.
Carbon↗
Microchip-based ethanol/oxygen biofuel cell.
One of the limitations of lab-on-a-chip technology has been the lack of integrated power supplies for powering various devices on the chip. This research focused on design of a stackable, microchip-based biofuel cell. The biofuel cell is powered by the addition of ethanol through a flow channel to a bioanode. The bioanode contains a micromolded carbon ink anode that has been modified with two layers. The first layer is poly(methylene green), which is an electrocatalyst for NADH oxidation; the second layer is a membrane that contains an immobilized enzyme, alcohol dehydrogenase. Each layer was characterized electrochemically. It was found that the poly(methylene green) layer is kinetically-limited, but when the complete bioanode is formed, the bioanode is diffusion-limited due to slow mass transport of NADH within the modified Nafion membrane. When used relative to an external platinum cathode, the biofuel cell showed maximum open circuit potentials of 0.34 V and maximum current densities of 53.0 +/- 9.1 microA cm(-2). This research demonstrates the feasibility of a microfabricated biofuel cell device.
Alcohol Dehydrogenase↗
Chembio extraction on a chip by nanoliter droplet ejection.
This paper describes a novel liquid separation technique for chembio extraction by an ultrasonic nanoliter-liquid-droplet ejector built on a PZT sheet. This technique extracts material from an aqueous two-phase system (ATPS) in a precise amount through digital control of the number of nanoliter droplets, without any mixing between the two liquids in the ATPS. The ultrasonic droplet ejector uses an acoustic streaming effect produced by an acoustic beam focused on the liquid surface, and ejects liquid droplets only from the liquid surface without disturbing most of the liquid below the surface. This unique characteristic of the focused acoustic beam is perfect (1) for separating a top-layer liquid (from the bulk of liquid) that contains particles of interest or (2) for recovering a top-layer liquid that has different phase from a bottom-layer liquid. Three kinds of liquid extraction are demonstrated with the ultrasonic droplet ejector: (1) 16 microl of top layer in Dextran-polyethylene glycol-water ATPS (aqueous two-phase system) is recovered within 20 s; (2) micron sized particles that float on water surface are ejected out with water droplets; and (3) oil layer on top of water is separated out.
Acoustics↗
Application of microchip assay system for the measurement of C-reactive protein in human saliva.
In the last decade, saliva has been advocated as a non-invasive alternative to blood as a diagnostic fluid. However, use of saliva has been hindered by the inadequate sensitivity of current methods to detect the lower salivary concentrations of many constituents compared to serum. Furthermore, developments in the areas related to lab-on-a-chip systems for saliva-based point of care diagnostics are complicated by the high viscosity and heterogeneous properties associated with this diagnostic fluid. The biomarker C-reactive protein (CRP) is an acute phase reactant and a well-accepted indicator of inflammation. Numerous clinical studies have established elevated serum CRP as a strong, independent risk factor for the development of cardiovascular disease (CVD). CVD has also been associated with oral infections (i.e. periodontal diseases) and there is evidence that systemic CRP may be a link between the two. Clinical measurements of CRP in serum are currently performed with "high sensitivity" CRP (hsCRP) enzyme-linked immunosorbent assay (ELISA) tests that lack the sensitivity for the detection of this important biomarker in saliva. Because measurement of salivary CRP may represent a novel approach for diagnosing and monitoring chronic inflammatory disease, including CVD and periodontal diseases, the objective of this study was to apply an ultra-sensitive microchip assay system for the measurement of CRP in human saliva. Here, we describe this novel lab-on-a-chip system in its first application for the measurement of CRP in saliva and demonstrate its advantages over the traditional ELISA method. The increased sensitivity of the microchip system (10 pg ml(-1) of CRP with 1000-fold dilution of saliva sample) is attributed to its inherent increased signal to noise ratio, resulting from the higher bead surface area available for antigen/antibody interactions and the high stringency washes associated with this approach. Finally, the microchip assay system was utilized in this study to provide direct experimental evidence that chronic periodontal disease may be associated with higher levels of salivary CRP.
C-Reactive Protein↗
Measuring reaction kinetics in a lab-on-a-chip by microcoil NMR.
A microfluidic chip with an integrated planar microcoil was developed for Nuclear Magnetic Resonance (NMR) spectroscopy on samples with volumes of less than a microliter. Real-time monitoring of imine formation from benzaldehyde and aniline in the microreactor chip by NMR was demonstrated. The reaction times in the chip can be set from 30 min down to ca. 2 s, the latter being the mixing time in the microfluidic chip. Design rules will be described to optimize the microreactor and detection coil in order to deal with the inherent sensitivity of NMR and to minimize magnetic field inhomogeneities and obtain sufficient spectral resolution.
Imines↗
Immobilization of individual cells by local photo-polymerization on a chip.
A novel separation method for random screening of target cells from a large heterogeneous population by using a local photo-polymerization is developed. A photo-crosslinkable resin solution is mixed with the sample liquid and we controlled the state from sol to gel by irradiating the near ultraviolet (UV) light with the mercury lamp and He-Cd laser near the target cell. We applied three types of immobilization methods such as direct immobilization method, caging method, and direct immobilization with position control method. The selected cell is immobilized in the cured resin directly or inside the cage of the cured resin. In the position control method, laser tweezers are employed to manipulate the target cell indirectly by using the droplet of the resin as a microtool. The cell is positioned properly by the laser manipulation system and is immobilized in the polymerized resin. After the selected cells are immobilized we can easily remove the other objects by the cleaning flow in the microchannel since the polymerized resin strongly binds with the cover glass and resists more than 466 mm s(-1) flow speed in the microchannel (microchannel size: width is 500 micron and depth is 100 micron). We tested the mercury lamp as well as the He-Cd laser for UV-light irradiation at the local area and confirmed improvement of resolution of the cured area by using the He-Cd laser (from 7 micron to 5 micron). Based on this method, we succeeded in single cell immobilization and basic experiments such as culture and fluorescent dyeing of immobilized yeast cells.
Cell Separation↗
Analysis of natural flavonoids by microchip-micellar electrokinetic chromatography with pulsed amperometric detection.
Catechins (catechin and other derivatives) are naturally occurring flavonoids present in a number of plants and foods. They are also part of numerous nutraceutical formulations because they are believed to have antioxidant, cancer chemo-preventative, anti-inflammatory and antimicrobial properties. The determination of catechins has traditionally been performed by HPLC. However, this methodology is both time and sample intensive and generates large amounts of organic solvent waste. In the current report, an application of MEKC using a PDMS microchip is presented for the analysis of catechins. The system uses pulsed amperometric detection for direct analysis of important naturally occurring catechins. The effect of pH, surfactant concentration, detection potential and signal stability were analyzed. Linear relationships were found between the concentration and peak current, with good stability and limits of detection of 8 [micro sign]M for catechin, epigallocatechin gallate and epicatechin, and 14 [micro sign]M for epicatechin gallate. Optimum conditions were applied to the detection of selected catechins in a commercially available green tea extract nutraceutical and the results were compared to HPLC analysis. The analysis using microchip micellar electrokinetic chromatography and pulsed amperometric detection was completed in 4.5 min, 10 times faster than the HPLC analysis.
Catechin↗
A MALDI-chip integrated system with a monitoring window.
The integration of a monitoring port along the microfluidic path of a MALDI-chip integrated device is described. Optimization of the microreactor design allows longer reaction and measuring times. The Schiff base reaction between 4-tert-butylaniline (1) and 4-tert-butylbenzaldehyde (2) in ethanol was carried out on-chip in the MALDI ionization chamber and the formed imine 3 was detected in real time, demonstrating the feasibility of the "monitoring window" approach. This preliminary result opens the way to on-chip kinetic studies by MALDI-MS, by opening multiple monitoring windows along the microchannel.
Aniline Compounds↗
Self-referencing a single waveguide grating sensor in a micron-sized deep flow chamber for label-free biomolecular binding assays.
In order to allow the design of increasingly sensitive label-free biosensors, compensation of environmental fluctuations is emerging as the dominant hurdle. The system and technique presented here utilize a unique combination of microfluidics, optical instrumentation, and image processing to provide a reference signal for each label-free biomolecular binding assay. Moreover, this reference signal is generated from the same sensor used to detect the biomolecular binding events. In this manner, the reference signal and the binding signal share nearly all common-mode noise sources (temperature, pressure, vibration, etc.) and their subtraction leaves the purest binding signal possible. Computational fluid dynamic simulations have been used to validate the flow behavior and thermal characteristics of the fluids inside the sensing region. This system has been demonstrated in simple bulk refractive index tests, as well as small molecule (biotin/streptavidin) binding experiments. The ability to perform not only simple binding but also control experiments has been discussed, indicating the wide applicability of the technique.
Biotin↗