Monitoring cellular stress responses to nanoparticles using a lab-on-a-chip

As nanotechnology moves towards widespread commercialization, new technologies are needed to adequately address the potential health impact of nanoparticles (NPs). Assessing the safety of over 30,000 NPs through animal testing would not only be expensive, but it would also raise a number of ethical considerations. Furthermore, existing in vitro cell-based assays are not sufficient in scope to adequately address the complexity of cell-nanoparticle interactions including NP translocation, accumulation and co-transport of e.g. allergens. In particular, classical optical/fluorescent endpoint detection methods are known to provide irreproducible, inaccurate and unreliable results since these labels can directly react with the highly catalytic surfaces of NP. To bridge this technological gap we have developed a lab-on-a-chip capable of continuously and non-invasively monitoring the collagen production of primary human fibroblast cells (NHDF) using contactless dielectric microsensors. Human dermal fibroblast cells are responsible for the maintenance of soft tissue integrity, are found throughout the human body and their primary function is collagen expression. We show that cellular collagen production can be readily detected and used to assess cellular stress responses to a variety of external stimuli, including exposure to nanoparticles. Results of the study showed a 20% and 95% reduction of collagen production following 4 hour exposure to 10 μg mL(-1) gold and silver nanoparticles (dia.10 nm), respectively. Furthermore a prolonged perfusion of sub-toxic concentrations (0.1 μg mL(-1)) of silver NP reduced NHDF collagen production by 40% after 10 h indicating increased NP take up and accumulation. We demonstrate that the application of microfluidics for the tailored administration of different NP treatments constitutes a powerful new tool to study cell-nanoparticle interactions and nanoparticle accumulation effects in small cell populations.     

Supravital fluorometric apoptosis detection in a single mouse embryo using lab-on-a-chip

Detection of apoptosis is one of the main criteria of preimplantation embryo growth potential assessment. Recent developments in lab-on-a-chip techniques has led to apoptosis detection and monitoring on a single cell or embryo level. However, single embryo apoptosis detection without a change in embryo developmental competence and post-examination "recovery" still remains a challenge. In this paper we present a lab-on-a-chip, co-working with miniaturized optical instrumentation, which allows supravital examination of single embryos for the presence of apoptotic blastomers with full after lab-on-a-chip study "recovery" and maintenance of their further developmental capacity.     

Rapid assessment of soil pollution with kerosene using a carbon-nanotube-based piezosensor

The possibility of determining the kerosene content in the ground using a flash detector with open cell detection based on a piezosensor is considered. The coating of the electrodes of a quartz crystal resonator and its weight for the detection of kerosene vapors in a wide concentration range, the structural parameters of the flash detector housing, the conditions for detecting kerosene vapors above ground are optimized. The accuracy and reliability of assessing of the level of kerosene contamination in different types of soils are estimated in field conditions.     

Rapid chromite dissolution using manganese dioxide-lithium-sulphate-sulphuric acid mixture-estimation of alumina in chromite for quality control assessment

Proposed in this paper is a less labour-intensive method involving a mixture (0.3 g MnO₂ + 1.0 g Li₂SO₄ + 10 mL H₂O + 20 mL of 36M H₂SO₄ for 0.2 g sample) to decompose chromites for accurate and precise estimation of alumina in chromite matrices for quality control (QC) assessment. Several international chromite samples, namely, CHR-BKg, CHR-Pt, IGS-30 and BCS-308 were analysed. Intermethod comparison studies reveal that the probability of results being different is less than (slight variation in case of CHR-BKg) 99% when two methods [ethylenediamine tetraacetic acid (EDTA) titration and spectrophotometry using ammonium aurinitricarboxylate (aluminon)] were compared. It also shows that the proposed method yields better results than the most conventional route involving HClO₄ decomposition used for similar purpose; hence can be used for QC programme and in the evaluation of reference materials.     

Rapid quality assessment of Radix Aconiti Preparata using direct analysis in real time mass spectrometry

This study presents a novel and rapid method to identify chemical markers for the quality control of Radix Aconiti Preparata, a world widely used traditional herbal medicine. In the method, the samples with a fast extraction procedure were analyzed using direct analysis in real time mass spectrometry (DART MS) combined with multivariate data analysis. At present, the quality assessment approach of Radix Aconiti Preparata was based on the two processing methods recorded in Chinese Pharmacopoeia for the purpose of reducing the toxicity of Radix Aconiti and ensuring its clinical therapeutic efficacy. In order to ensure the safety and effectivity in clinical use, the processing degree of Radix Aconiti should be well controlled and assessed. In the paper, hierarchical cluster analysis and principal component analysis were performed to evaluate the DART MS data of Radix Aconiti Preparata samples in different processing times. The results showed that the well processed Radix Aconiti Preparata, unqualified processed and the raw Radix Aconiti could be clustered reasonably corresponding to their constituents. The loading plot shows that the main chemical markers having the most influence on the discrimination amongst the qualified and unqualified samples were mainly some monoester diterpenoid aconitines and diester diterpenoid aconitines, i.e. benzoylmesaconine, hypaconitine, mesaconitine, neoline, benzoylhypaconine, benzoylaconine, fuziline, aconitine and 10-OH-mesaconitine. The established DART MS approach in combination with multivariate data analysis provides a very flexible and reliable method for quality assessment of toxic herbal medicine.     

Chemical and biological threat-agent detection using electrophoresis-based lab-on-a-chip devices

The ability to separate complex mixtures of analytes has made capillary electrophoresis (CE) a powerful analytical tool since its modern configuration was first introduced over 25 years ago. The technique found new utility with its application to the microfluidics based lab-on-a-chip platform (i.e., microchip), which resulted in ever smaller footprints, sample volumes, and analysis times. These features, coupled with the technique's potential for portability, have prompted recent interest in the development of novel analyzers for chemical and biological threat agents. This article will comment on three main areas of microchip CE as applied to the separation and detection of threat agents: detection techniques and their corresponding limits of detection, sampling protocol and preparation time, and system portability. These three areas typify the broad utility of lab-on-a-chip for meeting critical, present-day security, in addition to illustrating areas wherein advances are necessary.     

Rapid dissolution procedure for base-metals-bearing complex materials for quality control assessment

A less labour-intensive method, involving a mixture of 1.0 g MnO₂ + 20 mL HCl (11), is proposed for decomposing seven ores and six metallurgical products for accurate and precise estimation of copper, lead, zinc, cobalt and nickel for quality control assessment. Twentyone international reference materials were also analysed. Results of regression analyses are presented and intermethod comparison studies reveal that the probability of results being different compared with a reference method was less than 1%. Sample decomposition is straightforward and the method has been found to be very simple, rapid and easily adaptable, as it involves no separation of the analyte from the matrix elements.     

Compact fluorescence detection using in-fiber microchannels-its potential for lab-on-a-chip applications

This paper reports a compact and practical fluorescence sensor using an in-fiber microchannel. A blue LED, a multimode PMMA or silica fiber and a mini-PMT were used as an excitation source, a light guide and a fluorescence detector, respectively. Microfluidic channels of 100 microm width and 210 microm depth were fabricated in the optical fibers using a direct-write CO(2) laser system. The experimental results show that the sensor has high sensitivity, able to detect 0.005 microg L(-1) of fluorescein in the PBS solution, and the results are reproducible. The results also show that the silica fiber sensor has better sensitivity than that of the PMMA fiber sensor. This could be due to the fouling effect of the frosty layer formed at the microchannel made within the PMMA fiber. It is believed that this fiber sensor has the potential to be integrated into microfluidic chips for lab-on-a-chip applications.     

Drop mixing in a microchannel for lab-on-a-chip platforms

We present theory, simulations, and experiments for discrete drop mixing in microchannels. The drops are placed sequentially in a channel and then moved at a set velocity to achieve mixing. The mixing occurs in three different regimes (diffusion-dominated, dispersion-dominated, and convection-dominated) depending on the Péclet number (Pe) and the drop dimensions. Introducing the modified Péclet number (Pe*), we show asymptotic curves that can be used to predict the mixing time and the required distance for mixing for any of the three regimes. Simulations of the mixing experiments using COMSOL agree with the theoretical limits. In our experimental work, we used a polydimethylsiloxane (PDMS) microchannel with a membrane air bypass valve to remove the air between drops. This approach enables precise control of the mixing and merging site. Experimental, simulation, and theoretical results all agree and show that mixing can occur in fractions of a second to hours, depending on the parameters used.     

Rapid assessment of DNA damage induced by polystyrene nanosphere suspension using a photoelectrochemical DNA sensor

Nanomaterials have been used increasingly in a wide variety of applications, and some of them have shown toxic effects on experimental animals and cells. In this study, a previously established photoelectrochemical DNA sensor was employed to rapidly detect DNA damage induced by polystyrene nanosphere (PSNS) suspensions. In the sensor, a double-stranded DNA film was assembled on a semiconductor electrode, and a DNA intercalator, Ru(bpy)2(dppz)2+ (bpy = 2,2'-bipyridine, dppz = dipyrido[3,2-a:2',3'-c]phenazine...     

Isolation of plasma from whole blood using planar microfilters for lab-on-a-chip applications

Researchers are actively developing devices for the microanalysis of complex fluids, such as blood. These devices have the potential to revolutionize biological analysis in a manner parallel to the computer chip by providing very high throughput screening of complex samples and massively parallel bioanalytical capabilities. A necessary step performed in clinical chemistry is the isolation of plasma from whole blood, and effective sample preparation techniques are needed for the development of miniaturized clinical diagnostic devices. This study demonstrates the use of passive, operating entirely on capillary action, transverse-flow microfilter devices for the microfluidic isolation of plasma from whole blood. Using these planar microfilters, blood can be controllably fractionated with minimal cell lysis. A characterization of the device performance reveals that plasma filter flux is dependent upon the wall shear rate of blood in the filtration channel, and this result is consistent with macroscale blood filtration using microporous membranes. Also, an innovative microfluidic layout is demonstrated that extends device operation time via capillary action from seconds to minutes. Efficiency of these microfilters is approximately three times higher than the separation efficiencies predicted for microporous membranes under similar conditions. As such, the application of the microscale blood filtration designs used in this study may have broad implications in the design of lab-on-a-chip devices, as well as the field of separation science.     

Quantitative 3-dimensional profiling of channel networks within transparent lab-on-a-chip microreactors using a digital imaging method

We have developed a method for the quantitative 3-dimensional profiling of micron sized channel networks within optically transparent "lab-on-a-chip" microreactor devices. The method involves capturing digitised microscope images of the channel network filled with an optically absorbing dye. The microscope is operated in transmission mode using light filtered through a narrow bandpass filter with a maximum transmission wavelength matching the wavelength of the absorbance maximum of the dye solution. Digitised images of a chip filled with solvent and dye solution are analysed pixel by pixel to yield a spatially resolved array of absorbance values. This array is then converted to optical path length values using the Beer-Lambert law, thereby providing the 3D profile of the channel network. The method is capable of measuring channel depths from 10 to 500 microm (and probably even smaller depths) with an accuracy of a few percent. Lateral spatial resolution of less than 1 microm is achievable. It has been established that distortion of the measured profiles resulting from a mismatch in refractive index between the dye solution and the glass of the microreactors is insignificant. The method has been successfully used here to investigate the effects of thermal bonding and etch time on channel profiles. The technique provides a convenient, accurate and non-destructive method required to determine channel profiles; information which is essential to enable optimisation of the operating characteristics of microreactor devices for particular applications.     

A functional on-chip pressure generator using solid chemical propellant for disposable lab-on-a-chip

This paper presents a functional on-chip pressure generator that utilizes chemical energy from a solid chemical propellant to perform fluidic delivery in applications of plastic-based disposable biochips or lab-on-a-chip systems. In this functional on-chip pressure generator, azobis-isobutyronitrile (AIBN) as the solid chemical propellant is deposited on a microheater using a screen-printing technique, which can heat the AIBN at 70 degrees C to produce nitrogen gas. The output pressure of nitrogen gas, generated from the solid chemical propellant, is adjustable to a desired pressure by controlling the input power of the heater. Using this chemical energy source, the generated pressure depends on the deposited amount of the solid chemical propellant and the temperature of the microheater. Experimental measurements show that this functional on-chip pressure generator can achieve around 3 000 Pa pressure when 189 mJ of energy is applied to heat the 100 microg of AIBN. This pressure can drive 50 nl of water through a microfluidic channel of 70 mm and cross-sectional area of 100 microm x 50 microm. Due to its compact size, ease of fabrication and integration, high reliability (no moving parts), biologically inert gas output along with functionality of gas generation, this pressure generator will be an excellent pressure source for handling the fluids of disposable lab-on-a-chip, biochemical analysis systems or drug delivery systems.     

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.     

Rapid microwave-assisted extraction and HPTLC-photodensitometric method for the quality assessment of Boerhaavia diffusa L

A rapid and cost-effective method for the extraction of rotenoids in Boerhaavia diffusa L., based on the use of microwave-assisted extraction (MAE), is proposed. The conventional reflux, soxhlet, and maceration extraction methods were also conducted to validate the reliability of the new method. Under the optimized conditions, two rotenoids (boeravinone B and E) were extracted and quantified by HPTLC. The yield of boeravinone B and E achieved by MAE was 0.15 and 0.32% (w/w), respectively. The result showed that MAE-HPTLC is a simple, rapid, and solvent-sparing method for the extraction and quantitation of boeravinone B and E from B. diffusa L.     

Rapid quality assessment of Gentianae Macrophyllae Radix based on near-infrared spectroscopy and capillary electrophoresis

The aim of this study was to establish a rapid quality assessment method for Gentianae Macrophyllae Radix (RGM) using near-infrared (NIR) spectra combined with chemometric analysis. The NIR spectra were acquired using an integrating sphere diffuse reflectance module, using air as the reference. Capillary electrophoresis (CE) analyses were performed on a model P/ACE MDQ Plus system. Partial least squares-discriminant analysis qualitative model was developed to distinguish different species of RGM samples, and the prediction accuracy for all samples was 91%. The CE response values at each retention time were predicted by building a partial least squares regression (PLSR) calibration model with the CE data set as the Y matrix and the NIR spectra data set as the X matrix. The converted CE fingerprints basically match the real ones, and the six main peaks can be accurately predicted. Transforming NIR spectra fingerprints into the form of CE fingerprints increases its interpretability and more intuitively demonstrates the components that cause diversity among samples of different species and origins. Loganic acid, gentiopicroside, and roburic acid were considered quality indicators of RGM and calibration models were built using PLSR algorithm. The developed models gave root mean square error of prediction of 0.2592% for loganic acid, 0.5341% for gentiopicroside, and 0.0846% for roburic acid. The overall results demonstrate that the rapid quality assessment system can be used for quality control of RGM.     

Optical classification of algae species with a glass lab-on-a-chip

The identification of submillimetre phytoplankton is important for monitoring environmental and climate changes, as well as evaluating water for health reasons. Current standard methods for phytoplankton species identification require sample collection and ex situ analysis, an expensive procedure which prevents the rapid identification of phytoplankton outbreaks. To address this, we use a glass-based microchip with a microchannel and waveguide included on a monolithic substrate, and demonstrate its use for identifying phytoplankton species. The microchannel and the specimens inside it are illuminated by laser light from the curved waveguide as algae-laden water is passed through the channel. The intensity distribution of the light collected from the biochip is monitored with an external photodetector. Here, we demonstrate that the characteristics of the photodiode signal from this simple and robust system can provide significant and useful information as to the contents of the channel. Specifically, we show first that the signals are correlated to the size of algae cells. Using a pattern-matching neural network, we demonstrate the successful classification of five algae species with an average 78% positive identification rate. Furthermore, as a proof-of-concept for field-operation, we show that the chip can be used to distinguish between detritus in field-collected water and the toxin-producing cyanobacterium Cyanothece.     

Microfluidic designs and techniques using lab-on-a-chip devices for pathogen detection for point-of-care diagnostics

Effective pathogen detection is an essential prerequisite for the prevention and treatment of infectious diseases. Despite recent advances in biosensors, infectious diseases remain a major cause of illnesses and mortality throughout the world. For instance in developing countries, infectious diseases account for over half of the mortality rate. Pathogen detection platforms provide a fundamental tool in different fields including clinical diagnostics, pathology, drug discovery, clinical research, disease outbreaks, and food safety. Microfluidic lab-on-a-chip (LOC) devices offer many advantages for pathogen detection such as miniaturization, small sample volume, portability, rapid detection time and point-of-care diagnosis. This review paper outlines recent microfluidic based devices and LOC design strategies for pathogen detection with the main focus on the integration of different techniques that led to the development of sample-to-result devices. Several examples of recently developed devices are presented along with respective advantages and limitations of each design. Progresses made in biomarkers, sample preparation, amplification and fluid handling techniques using microfluidic platforms are also covered and strategies for multiplexing and high-throughput analysis, as well as point-of-care diagnosis, are discussed.     

High-throughput assay of nitric oxide metabolites in human plasma without deproteinization by lab-on-a-chip electrophoresis using a zwitterionic additive

In order to develop a high-throughput assay for nitric oxide metabolites, nitrite (NO2-) and nitrate (NO3-), in biological fluids, we have investigated the simultaneous determination of them using an electrophoretic lab-on-a-chip (microchip capillary electrophoresis, MCE) technique. In this study, in order to establish an MCE assay process without deproteinization, the addition of a zwitterionic additive into the running buffer to reduce the adsorption of protein onto the surface of channel was investigated. Initially, some zwitterionic additives were investigated by making a comparison of relative standard deviations (RSDs) of the migration times for NO2(-) and NO3(-) on capillary electrophoresis. From the results of our comparison of the RSD values, 2% (w/w) N-cyclohexyl-2-aminoethanesulfonic acid (CHES) was selected. As a result of the application of the running buffer with CHES to the MCE process, the complete separation of NO2(-) and NO3(-) in human plasma without deproteinization was achieved within 1 min. Since the RSD values of the positions of the peaks were less than 2.3%, beneficial reduction effects on MCE were suggested. When we used an internal standard method in order to correct the injection volume, the RSDs of the peak heights and areas were less than 10%, and the correlation coefficients of spiked calibration curves ranging from 0 to 350 microM were 0.999 and 0.997 for NO2(-) and NO3(-), respectively. The limits of detection (S/N=3) were 53 microM for NO2(-) and 41 microM for NO3(-). Moreover, the correlation coefficients in excess of 0.99 between the MCE method and a conventional Griess method were achieved for both NO2(-) and NO3(-). Consequently, the possibility of establishing a high-throughput assay process was obtained by utilizing 2% (w/w) CHES to reduce protein adsorption.