Recent advances in droplet microfluidics for microbiology

Advances in microbiology rely on innovations in technology. Droplet microfluidics, as a versatile and powerful technique that allows high-throughput generation and manipulation of subnanoliter volume droplets, has become an indispensable tool shifting experimental paradigms in microbiology. Droplet microfluidics has opened new avenues to various microbiological research, from resolving single-cell heterogeneity to investigating spatiotemporal dynamics of microbial communities, from precise quant...     

On-line octane-number analyser for reforming unit effluents. Principle of the analyser and test of a prototype

An on-line automatic explosion-proof process octane-number analyser was developed. The principle is based on on-line capillary gas chromatographic analysis of the sample. A computer program performs the identification and the quantification of the components and calculates the octane numbers and other physical properties. The process analyser consists of laboratory equipment placed in an explosion-proof shelter. A prototype of the analyser was tested on a pilot plant under industrial conditions. It was shown that the fully automated analyser is reliable and reproducible.     

A pyrometric differential thermal analyser for high temperatures

An apparatus for differential thermal analysis to a temperature of 3400°C is described. The specimen is heated inductively with a frequency of 1 MHz either without crucible at its upper end only or in a crucible which is one of the components of the system investigated. The differential principle is applied by comparing the radiation emitted from the sample with the power fed to the sample, which is registered by a bulb coupled to the high frequency circuit. The emission of the sample and bulb are measured by two photodiodes. The different behaviour of sample and bulb is compensated electronically. Investigations in metal ceramic eutectics are given as an example.     

Electrowetting-based droplet mixers for microfluidic systems

Mixing of analytes and reagents is a critical step in realizing a lab-on-a-chip. However, mixing of liquids is very difficult in continuous flow microfluidics due to laminar flow conditions. An alternative mixing strategy is presented based on the discretization of liquids into droplets and further manipulation of those droplets by electrowetting. The interfacial tensions of the droplets are controlled with the application of voltage. The droplets act as virtual mixing chambers, and mixing occurs by transporting the droplet across an electrode array. We also present an improved method for visualization of mixing where the top and side views of mixing are simultaneously observed. Microliters of liquid droplets are mixed in less than five seconds, which is an order of magnitude improvement in reported mixing times of droplets. Flow reversibility hinders the process of mixing during linear droplet motion. This mixing process is not physically confined and can be dynamically reconfigured to any location on the chip to improve the throughput of the lab-on-a-chip.     

Principles of droplet electrohydrodynamics for lab-on-a-chip

Electrically controlled droplet-based labs-on-a-chip operate under the principles of electro-capillarity and dielectrophoresis. The microfluidic mechanics of manipulating electrified droplets are complex and not entirely understood. In this article, we analyse these operating principles, especially electrowetting on dielectric (a form of electro-capillarity) and dielectrophoresis, under a unified framework of droplet electrohydrodynamics. We differentiate them by their electric origins and their energy transduction mechanisms. Our study shows that both electrowetting on dielectric and dielectrophoresis are effective for droplet generation and manipulation. In addition, our study demonstrates: (1) the presence of a wetting contribution to dielectrophoresis; and (2) contact angle reduction is merely an observable consequence of, not a condition for, the occurrence of electrowetting on dielectric. Simulations are used extensively in this article to illustrate device operation, to expose underlying physics, and to validate our conclusions. Simulations of electrically driven droplet generation, droplet translocation, droplet fusion, and droplet fission are presented.     

Alternating droplet generation and controlled dynamic droplet fusion in microfluidic device for CdS nanoparticle synthesis

A multifunctional and high-efficiency microfluidic device for droplet generation and fusion is presented. Through unique design of the micro-channels, the device is able to alternately generate droplets, generating droplet ratios ranging from 1 ratio 5 to 5 ratio 1, and fuse droplets, enabling precise chemical reactions in several picoliters on a single chip. The controlled fusion is managed by passive control based on the channel geometry and liquid phase flow. The synthesis of CdS nanoparticles utilizing each fused droplet as a microreactor for rapid and efficient mixing of reagents is demonstrated in this paper. Following alternating droplet generation, the channel geometry allows the exclusive fusion of alternate droplets with concomitant rapid mixing and produces supersaturated solution of Cd2+ and S2- ions to form CdS nanoparticles in each fused droplet. The spectroscopic properties of the CdS nanoparticles produced by this method are compared with CdS prepared by bulk mixing.     

A new electrochemical analyser for nitric oxide and nitrogen dioxide

Individual sensors employing Teflon-bonded diffusion electrodes have been developed to measure nitric oxide and nitrogen dioxide separately, simultaneously, and continuously at part-permillion levels. The NO sensor was biased at 1.5V and that for NO₂ at 0.8V, both relative to the hydrogen electrode. The crucial factor in the virtual elimination of response from carbon monoxide, a relatively abundant air-pollutant, was the use of gold electrodes for both detectors. At 0.8 V NO does not react on gold. Although NO₂ does oxidize at 1.5 V it was removed quantitatively from NO/NO₂ mixtures by triethanolamine on firebrick. The NO₂ reduction signal and the NO oxidation signal were stable and reproducible.     

Water droplet bouncing--a definition for superhydrophobic surfaces

The ability of water to bounce on a surface provides an indication of many of the surface's properties. The technique described in this article uses water bouncing to determine the hydrophobicity of a surface, with a relationship established between water contact angle and number of bounces, which is dependent on the surfaces microstructure.     

Rapid droplet mixers for digital microfluidic systems

The mixing of analytes and reagents for a biological or chemical lab-on-a-chip is an important, yet difficult, microfluidic operation. As volumes approach the sub-nanoliter regime, the mixing of liquids is hindered by laminar flow conditions. An electrowetting-based linear-array droplet mixer has previously been reported. However, fixed geometric parameters and the presence of flow reversibility have prevented even faster droplet mixing times. In this paper, we study the effects of varying droplet aspect ratios (height:diameter) on linear-array droplet mixers, and propose mixing strategies applicable for both high and low aspect ratio systems. An optimal aspect ratio for four electrode linear-array mixing was found to be 0.4, with a mixing time of 4.6 seconds. Mixing times were further reduced at this ratio to less than three seconds using a two-dimensional array mixer, which eliminates the effects of flow reversibility. For lower aspect ratio (相似文献   

French laboratory proficiency testing program for food microbiology

The proficiency testing program in food microbiology (Réseau d'Analyses et d'Echanges en Microbiologie des Aliments; RAEMA), created in 1988, currently includes 440 participating laboratories. The program establishes proficiency in detection of Salmonella and Listeria monocytogenes, as well as quantitation of aerobic microorganisms, Enterobacteriaceae, coliforms, Escherichia coli, Clostridium perfringens, coagulase-positive Staphylococcus, and Listeria monocytogenes. Twice a year, 5 test samples are sent to participants to assess their precision and trueness for enumeration and detection of microorganisms. Results show an increasing involvement of food microbiology laboratories in quality assurance programs and use of standard and validated analytical methods. However, the percentage of laboratories obtaining questionable and unsatisfactory microbiological results remains relatively constant.     

Simplified low-pressure high-resolution nucleotide analyser.

A simple nucleotide analyser was constructed from commercially available components. A 20 cm X 0.4 cm I.D. column of pellicular anion exchanger operating at approximately 20 p.s.i. was found to perform identically with published methods using high-pressure equipment. The eluant was monitored using a variable-wavelength UV monitor. A very simple device for producing variable gradients was employed. The nucleotide analyser was trouble free to operate and the results were both linear and reproducible. Applications to the rapid analysis of nucleotides and the high-sensitivity analysis (approx. 10 pmoles) of cyclic 3',5'-adenosine monophosphate and cyclic 2',3'-guanosine monophosphate and tissue nucleotides are described.     

Self-aligned wet-cell for hydrated microbiology observation in TEM

This paper describes a Self-Aligned Wet (SAW) cell suitable for direct-cell or bacteria incubation and observation in a wet environment inside a transmission electron microscope. This SAW cell is fabricated by a bulk-micromachining process and composed of two structurally complementary counterparts (an out-frame and an in-frame), where each contain a silicon nitride film based observation window. The in- and out-frames can be self-aligned via a mechanism of surface tension from a bio-sample droplet without the aid of positioning stages. The liquid chamber is enclosed between two silicon nitride membranes that are thin enough to allow high energy electrons to penetrate while also sustaining the pressure difference between the TEM vacuum and the vapor pressure within the liquid chamber. A large field of view (150 μm × 150 μm) in a SAW cell is favored and formed from a larger sized observation window in the out-frame, which is fabricated using a unique circular membrane formation process. In this paper, we introduce a novel design to circumvent the challenges of charging/heating problems in silicon nitride that arise from interactions with an electron beam. This paper also demonstrates TEM observations of D. Radiodurans growth in a liquid environment within a thicker chamber (20 μm) within a SAW cell.     

Stability of a NDIR analyser for CO2 at atmospheric concentration

Carbon dioxide monitoring is significant in the environmental field since this gas plays an important role in the greenhouse effect. In order to determine CO2 concentration and to develop simulation models, it is necessary to carry out measurements which are accurate and comparable in time and space, i.e. SI-traceable. Non-dispersive infrared (NDIR) analysers are employed for CO2 measurements, as they are precise and stable. In order to achieve traceability, such instruments have to be characterized and calibrated. At the Istituto di Metrologia "G. Colonnetti"--CNR, a procedure for calibrating NDIR analysers for CO2 at atmospheric level was developed, which enables to calculate a correction for the analyser output. In addition, a complete uncertainty analysis was carried out and a correct traceability chain was established. The goal of the present work is the study of the stability of a NDIR analyser by repeating calibrations during three years and comparing the correction curves obtained to identify a proper re-calibration interval for such analysers. The investigated instrument has good repeatability and reproducibility, hence satisfactory stability during time, as shown by the short-term and long-term compatibility of calibration curves.     

An integrated portable hand-held analyser for real-time isothermal nucleic acid amplification

A compact hand-held heated fluorometric instrument for performing real-time isothermal nucleic acid amplification and detection is described. The optoelectronic instrument combines a Printed Circuit Board/Micro Electro Mechanical Systems (PCB/MEMS) reaction detection/chamber containing an integrated resistive heater with attached miniature LED light source and photo-detector and a disposable glass waveguide capillary to enable a mini-fluorometer. The fluorometer is fabricated and assembled in planar geometry, rolled into a tubular format and packaged with custom control electronics to form the hand-held reactor. Positive or negative results for each reaction are displayed to the user using an LED interface. Reaction data is stored in FLASH memory for retrieval via an in-built USB connection. Operating on one disposable 3 V lithium battery >12, 60 min reactions can be performed. Maximum dimensions of the system are 150 mm (h) × 48 mm (d) × 40 mm (w), the total instrument weight (with battery) is 140 g. The system produces comparable results to laboratory instrumentation when performing a real-time nucleic acid sequence-based amplification (NASBA) reaction, and also displayed comparable precision, accuracy and resolution to laboratory-based real-time nucleic acid amplification instrumentation. A good linear response (R² = 0.948) to fluorescein gradients ranging from 0.5 to 10 μM was also obtained from the instrument indicating that it may be utilized for other fluorometric assays. This instrument enables an inexpensive, compact approach to in-field genetic screening, providing results comparable to laboratory equipment with rapid user feedback as to the status of the reaction.     

A microprocessor-controlled flow injection analyser for the determination of terbutaline sulphate

The determination is based on the reaction with 4-aminoantipyrine and potassium hexacyanoferrate(III). The system is automatic with a microcomputer controlling the sampler and injection valve, reading the output from the spectrophotometer and evaluating the result. The system is intended for drug analysis, and the matrix viscosity can vary considerably between different formulations without affecting peak area measurements.