Combined physico-chemical and biological sensing in environmental monitoring

Single-use conductivity and microbial sensors were used to investigate the effect of both species (chloride, nitrate, and sulphate) and concentration/osmolarity of anions on the metabolic activity of Escherichia coli. A new disposable, single-use conductivity sensor is described which is compatible with the CellSense mediated amperometric biosensor system. The effect of changing salinity and nitrate concentration on the response of E. coli to 3,5-dichlorophenol and mercuric chloride was determined. The implications for toxicity assessment of a hybrid sensing system, allowing the simultaneous monitoring of physico-chemical and biological data, are discussed.     

Lactic Acid Bacteria Biomass Monitoring in Highly Conductive Media by Permittivity Measurements

Recent technological improvements have extended the application range of permittivity biomass measurements to bacterial fermentations in highly conductive media. With Lactobacillus casei, the effective biomass detection sensitivity of the FOGALE Biomass System is around 0.2 g/l (0.01 pF/cm). Fermentations growth kinetics of Lactobacillus casei can be recorded with good reproducibility and accuracy despite the high medium conductivity varying between 15 and 75 mS/cm, and the low cell concentration (<6 g/l).     

Dielectric properties of yeast cells as determined by electrorotation.

Electrorotational spectra of yeast cells, Saccharomyces cerevisiae strain R XII, were measured over a frequency range of nearly 7 decades. The physical properties of distinct cell parts were simultaneously determined for individual cells by comparison with an electrical two-shell model: The conductivity of the cytoplasm, cell wall and cytoplasmic membrane of living cells were found to be 5.5 mS/cm, 0.1 to more than 0.5 mS/cm and less than 0.25 nS/cm to 4.5 microS/cm, respectively. The conductivity of the cytoplasmic membrane was dependent on the conductivity of the medium. Membrane behaviour is interpreted as an opening of membrane channels when the environment becomes more physiological. The specific membrane capacitance was determined to be 1.1 microF/cm2 and the thickness of the cell wall was calculated as 0.11 micron. Heat treated cells showed an increased membrane conductivity of more than 0.1 microS/cm (at 25 microS/cm medium conductivity) and a drop in cytoplasmic conductivity to between 0.1 and 0.8 mS/cm, depending on the length of time the cells were suspended in low conductivity water (25 microS/cm), indicating a perforation of the membrane. A slightly decreased spinning speed scaling factor for dead cells suggests a modification to the cellular surface, while the principal structure of the cell wall appears to be uneffected. It can be demonstrated by these observations, that cellular electrorotation permits the simultaneous investigation of the different cellular compartments of individual cells in vivo under various environmental conditions.     

Differences in membrane properties between unfertilised and fertilised single rabbit oocytes demonstrated by electro-rotation. Comparison with cells from early embryos

The apparent membrane capacity of tubular rabbit oocytes increases from 1.7-2.0 microF/cm2 before fertilisation to 3.7-4.0 microF/cm2 after fertilisation. The membrane conductivity measured on single cells was also increased by fertilisation from less than 1 mS/cm2 to 14 mS/cm2. Cells obtained from 2-, 4- or 8-cell embryos exhibited intermediate values of membrane capacity (2.3-2.8 microF/cm2) and conductivity (5-22 mS/cm2). The values quoted are those effective between 1 and 10 kHz, the frequency of the rotating field used. The large apparent capacities are probably due to the presence of structures such as microvilli which cause the actual membrane area to exceed the smooth sphere area. It must be assumed that these structures change in form or number on fertilisation, and that they persist in embryos, at least up to the 8-cell stage. No difference was apparent between cells fertilised in vitro or in vivo. Comparison of the above zona-free data with measurements on zona-complete oocytes indicate how fertilised and unfertilised rabbit eggs may be distinguished from one another, even in the presence of the zona pellucida.     

Ion exchange chromatography of antibody fragments

Effects of pH and conductivity on the ion exchange chromatographic purification of an antigen-binding antibody fragment (Fab) of pI 8.0 were investigated. Normal sulfopropyl (SP) group modified agarose particles (SP Sepharosetrade mark Fast Flow) and dextran modified particles (SP Sepharose XL) were studied. Chromatographic measurements including adsorption isotherms and dynamic breakthrough binding capacities, were complemented with laser scanning confocal microscopy. As expected static equilibrium and dynamic binding capacities were generally reduced by increasing mobile phase conductivity (1-25 mS/cm). However at pH 4 on SP Sepharose XL, Fab dynamic binding capacity increased from 130 to 160 (mg/mL media) as mobile phase conductivity changed from 1 to 5 mS/cm. Decreasing protein net charge by increasing pH from 4 to 5 at 1.3 mS/cm caused dynamic binding capacity to increase from 130 to 180 mg/mL. Confocal scanning laser microscopy studies indicate such increases were due to faster intra-particle mass transport and hence greater utilization of the media's available binding capacity. Such results are in agreement with recent studies related to ion exchange of whole antibody molecules under similar conditions.     

Electricity generation and treatment of paper recycling wastewater using a microbial fuel cell

Increased interest in sustainable agriculture and bio-based industries requires that we find more energy-efficient methods for treating cellulose-containing wastewaters. We examined the effectiveness of simultaneous electricity production and treatment of a paper recycling plant wastewater using microbial fuel cells. Treatment efficiency was limited by wastewater conductivity. When a 50 mM phosphate buffer solution (PBS, 5.9 mS/cm) was added to the wastewater, power densities reached 501 +/- 20 mW/m(2), with a coulombic efficiency of 16 +/- 2%. There was efficient removal of soluble organic matter, with 73 +/- 1% removed based on soluble chemical oxygen demand (SCOD) and only slightly greater total removal (76 +/- 4%) based on total COD (TCOD) over a 500-h batch cycle. Cellulose was nearly completely removed (96 +/- 1%) during treatment. Further increasing the conductivity (100 mM PBS) increased power to 672 +/- 27 mW/m(2). In contrast, only 144 +/- 7 mW/m(2) was produced using an unamended wastewater (0.8 mS/cm) with TCOD, SCOD, and cellulose removals of 29 +/- 1%, 51 +/- 2%, and 16 +/- 1% (350-h batch cycle). These results demonstrate limitations to treatment efficiencies with actual wastewaters caused by solution conductivity compared to laboratory experiments under more optimal conditions.     

A viscosity-dependent affinity sensor for continuous monitoring of glucose in biological fluids

We present a viscometric affinity biosensor for continuous monitoring of glucose in biological fluids such as blood and plasma. The sensing principle of this chemico-mechanical sensor is based upon the viscosity variation of a sensitive fluid with glucose concentration. Basically, this device includes both an actuating and a sensing piezoelectric diaphragms as well as a flow-resistive microchannel. In order to confine the sensitive fluid and allow glucose diffusion into the sensor, a free-standing alumina nanoporous membrane is also used as size-selective interface. Measurements carried out at nominal temperatures of 25 and 37 °C reveal that this sensor topology exhibits a high resolution in the current range of physiological blood glucose concentrations, i.e. 2-20 mM. In addition, complete reversibility was also demonstrated for at least 3 days. Finally, measurements performed in human blood serum confirm that this sensor fulfils all basic requirements for a use in continuous glucose monitoring of biological fluids.     

Use of inexpensive pressure transducers for measuring water levels in wells

Frequent measurement of below ground water levels at multiple locations is an important component of many wetland ecosystem studies. These measurements, however, are usually time consuming, labor intensive, and expensive. This paper describes a water-level sensor that is inexpensive and easy to construct. The sensor is placed below the expected low water level in a shallow well and, when connected to a datalogger, uses a pressure transducer to detect groundwater or surface water elevations. Details of pressure transducer theory, sensor construction, calibration, and examples of field installations are presented. Although the transducers must be individually calibrated, the sensors have a linear response to changing water levels (r ² .999). Measurement errors resulting from temperature fluctuations are shown to be about 4 cm over a 35°C temperature range, but are minimal when the sensors are installed in groundwater wells where temperatures are less variable. Greater accuracy may be obtained by incorporating water temperature data into the initial calibration (0.14 cm error over a 35C temperature range). Examples of the utility of these sensors in studies of groundwater/surface water interactions and the effects of water level fluctuations on tree growth are provided.     

Passive wireless MEMS microphones for biomedical applications

This paper introduces passive wireless telemetry based operation for high frequency acoustic sensors. The focus is on the development, fabrication, and evaluation of wireless, battery-less SAW-IDT MEMS microphones for biomedical applications. Due to the absence of batteries, the developed sensors are small and as a result of the batch manufacturing strategy are inexpensive which enables their utilization as disposable sensors. A pulse modulated surface acoustic wave interdigital transducer (SAW-IDT) based sensing strategy has been formulated. The sensing strategy relies on detecting the ac component of the acoustic pressure signal only and does not require calibration. The proposed sensing strategy has been successfully implemented on an in-house fabricated SAW-IDT sensor and a variable capacitor which mimics the impedance change of a capacitive microphone. Wireless telemetry distances of up to 5 centimeters have been achieved. A silicon MEMS microphone which will be used with the SAW-IDT device is being microfabricated and tested. The complete passive wireless sensor package will include the MEMS microphone wire-bonded on the SAW substrate and interrogated through an on-board antenna. This work on acoustic sensors breaks new ground by introducing high frequency (i.e., audio frequencies) sensor measurement utilizing SAW-IDT sensors. The developed sensors can be used for wireless monitoring of body sounds in a number of different applications, including monitoring breathing sounds in apnea patients, monitoring chest sounds after cardiac surgery, and for feedback sensing in compression (HFCC) vests used for respiratory ventilation. Another promising application is monitoring chest sounds in neonatal care units where the miniature sensors will minimize discomfort for the newborns.     

Effect of Electrical Conductivity and Silicate on Infection of Basil with Colletotrichum gloeosporioides in Soilless Culture

The aim of this study was to evaluate the effect of potassium silicate administration and of electrical conductivity of nutrient solution in three experiments against Colletotrichum gloeosporioides infection on basil (Ocimum basilicum L. cv Genovese Gigante) grown in a closed soilless system. Potassium silicate was added at 100 mg/l of nutrient solution at three different levels of electrical conductivity: 1.5–1.6 mS/cm (E.C.1), 3–3.2 mS/cm (E.C.2, 0.70 g/l NaCl) and 4–4.2 mS/cm (E.C.3, 0.95 g/l NaCl). Basil plants were inoculated with C. gloeosporioides spores 21–31 days after sowing or placing the pots on the channels, applying 5 ml of conidial suspension to each treatment. The increased electrical conductivity of the nutrient solution generally reduced the incidence and severity of the disease, with the highest electrical conductivity (E.C.3) providing the best results. The addition of potassium silicate to the different nutrient solutions showed a significant reduction in both incidence and severity of the disease compared to a solution without silicate and the best results were given by the addition of silicate with the highest electrical conductivity (E.C.3) in all the trials carried out. The combination of high electrical conductivity and potassium silicate supplied gave good results. The possibility and benefits of applying Si amendments in practice are examined.     

Continuous optical in-line glucose monitoring and control in CHO cultures contributes to enhanced metabolic efficiency while maintaining darbepoetin alfa product quality

Great efforts are directed towards improving productivity, consistency and quality of biopharmaceutical processes and products. One particular area is the development of new sensors for continuous monitoring of critical bioprocess parameters by using online or in-line monitoring systems. Recently, we developed a glucose biosensor applicable in single-use, in-line and long-term glucose monitoring in mammalian cell bioreactors. Now, we integrated this sensor in an automated glucose monitoring and feeding system capable of maintaining stable glucose levels, even at very low concentrations. We compared this fed-batch feedback system at both low (< 1 mM) and high (40 mM) glucose levels with traditional batch culture methods, focusing on glycosylation and glycation of the recombinant protein darbepoetin alfa (DPO) produced by a CHO cell line. We evaluated cell growth, metabolite and product concentration under different glucose feeding strategies and show that continuous feeding, even at low glucose levels, has no harmful effects on DPO quantity and quality. We conclude that our system is capable of tight glucose level control throughout extended bioprocesses and has the potential to improve performance where constant maintenance of glucose levels is critical.     

Thermal Stability of the Iron–Lactoferrin Complex in Aqueous Solution is Improved by Soluble Soybean Polysaccharide

Lactoferrin (Lf) can solubilize more than a 70-fold molar equivalent of iron in the presence of bicarbonate anions. The resulting iron?CLf complex (FeLf) is a useful food ingredient for iron fortification to prevent anemia. Although FeLf has greater thermal stability than Lf, a pasteurizing technique for FeLf has not been established. The aim of the present study was to develop a practical technique to pasteurize FeLf based on its thermal stability with the aid of a polysaccharide. FeLf [0.1?%, weight/weight (w/w) ratio] was heated at 80?°C for 3?min under various pH (5.5?C8.0) and electrical conductivity (0.1?C6.0?mS/cm) levels. Overall, FeLf was thermally stable and was hardly affected by pH or electrical conductivity, although aggregation and precipitation occurred when FeLf was heated at pH 6.0?C7.5 in the presence of salt and electrical conductivity >3.0?mS/cm. When 0.01?%?C0.4?% (w/w) of soluble soybean polysaccharide (SSPS) was added to 0.01?% (w/w) FeLf solution, the FeLf remained soluble and maintained its iron-holding property at pH 6.5, even when heated at 120?°C for 4?min. Particle charge measurements showed that the ??-potential of FeLf-SSPS became negatively charged following the addition of SSPS, which was associated with the improved thermal stability of FeLf. These results have important implications for the use of FeLf in developing liquid-based food products.     

Graphene–Polymer Nanocomposite‐Based Redox‐Induced Electricity for Flexible Self‐Powered Strain Sensors

Graphene as a powerful inorganic material such as excellent conductivity and ideal mechanical strength has recently been extensively utilized to develop flexible strain sensors. However, graphene‐based strain sensors usually suffer from the deficiencies of stretchability, sensitivity, and sensing range, which can restrict their applications in wearable devices. Here, a novel strain sensor is designed by integrating graphene/ecoflex film and meandering zinc wire into the flexible base. The constructed strain sensor not only possesses high stretchability of up to 150% strain but can also self‐generates current signals from redox‐induced electricity, where the stable current and voltage signals of about 75 µA and 0.83 V can be obtained, respectively. Furthermore, the self‐powered sensor presents a broad and linear sensing range of 25% to 150% strains and a fast response time of less than 0.11 s. Attached on human body, the sensor has been utilized to realize the motion detection of knee joint.     

Bimetallic PtM (M=Pd, Ir) nanoparticle decorated multi-walled carbon nanotube enzyme-free, mediator-less amperometric sensor for H₂O₂

A new highly catalytic and intensely sensitive amperometric sensor based on PtM (where M=Pd, Ir) bimetallic nanoparticles (NPs) for the rapid and accurate estimation of hydrogen peroxide (H(2)O(2)) by electrooxidation in physiological conditions is reported. PtPd and PtIr NPs-decorated multiwalled carbon nanotube nanocatalysts (PtM/MWCNTs) were prepared by a modified Watanabe method, and were characterized by XRD, TEM, ICP, and XAS. The sensors were constructed by immobilizing PtM/MWCNTs nanocatalysts in a Nafion film on a glassy carbon electrode. Both PtPd/MWCNTs and PtIr/MWCNTs assemblies catalyzed the electrochemical oxidation of H(2)O(2). Cyclic voltammetry characterization measurements revealed that both the PtM (M=Pd, Ir)/MWCNTs/GCE possessed similar electrochemical surface areas (～0.55 cm(2)), and electron transfer rate constants (～1.23 × 10(-3)cms(-1)); however, the PtPd sensor showed a better performance in H(2)O(2) sensing than did the PtIr counterpart. Explanations were sought from XAS measurements to explain the reasons for differences in sensor activity. When applied to the electrochemical detection of H(2)O(2), the PtPd/MWCNTs/GC electrode exhibited a low detection limit of 1.2 μM with a wide linear range of 2.5-125 μM (R(2)=0.9996). A low working potential (0V (SCE)), fast amperometric response (<5s), and high sensitivity (414.8 μA mM(-1)cm(-2)) were achieved at the PtPd/MWCNTs/GC electrode. In addition, the PtPd/MWCNTs nanocatalyst sensor electrode also exhibited excellent reproducibility and stability. Along with these attractive features, the sensor electrode also displayed very high specificity to H(2)O(2) with complete elimination of interference from UA, AA, AAP and glucose.     

Analysis of the sensitivity and frequency characteristics of coplanar electrical cell-substrate impedance sensors

A PDMS-glass based micro-device was designed and fabricated with 12 coplanar impedance sensors integrated for electrical cell-substrate impedance sensing (ECIS). The sensitivity and frequency characteristics of the sensors were investigated both theoretically (equivalent circuit model) and experimentally for the commonly used micro-electrode dimension scale (20-80 microm). The experimental results matched well with the theoretical model analysis and revealed that, within this micro-electrode dimension scale, as the electrode width decreased or as the total electrode length decreased the sensitivity of sensor increased over the whole sensing frequency range, whilst electrode to electrode distance had no influence on sensitivity. Through our frequency characteristics analysis, the whole frequency range could be divided into four parts. New functions describing the dominant components in each frequency range were defined and validated experimentally, and could be used to explain the phenomenon of an ECIS sensing frequency window. The contribution to the impedance measurement of cells growing on the edges of the electrodes was determined for the first time. Finally, novel proposals for ECIS sensor design and ECIS measurements were presented.     

Importance of temperature and anodic medium composition on microbial fuel cell (MFC) performance

The performance of a microbial fuel cell (MFC) was investigated at different temperatures and anodic media. A lag phase of 30 h occurred at 30 degrees C which was half that at room temperature (22 degrees C). The maximum power density at 30 degrees C was 70 mW/m(2) and at 22 degrees C was 43 mW/m(2). At 15 degrees C, no successful operation was observed even after several loadings for a long period of operation. Maximum power density of 320 mW/m(2) was obtained with wastewater medium containing phosphate buffer (conductivity: 11.8 mS/cm), which was approx. 4 times higher than the value without phosphate additions (2.89 mS/cm).     

Comparisons of optical pH and dissolved oxygen sensors with traditional electrochemical probes during mammalian cell culture

Small-scale upstream bioprocess development often occurs in flasks and multi-well plates. These culturing platforms are often not equipped to accurately monitor and control critical process parameters; thus they may not yield conditions representative of manufacturing. In response, we and others have developed optical sensors that enable small-scale process monitoring. Here we have compared two parameters critical to control in industrial cell culture, pH and dissolved oxygen (DO), measured with our optical sensors versus industrially accepted electrochemical probes. For both optical sensors, agreement with the corresponding electrochemical probe was excellent. The Pearson Correlations between the optical sensors and electrochemical probes were 98.7% and 99.7%, for DO and pH, respectively. Also, we have compared optical pH sensor performance in regular (320 mOsm/kg) and high-osmolality (450 mOsm/kg) cell culture media to simulate the increase in osmolality in pH-controlled cultures. Over a pH range of 6.38-7.98 the average difference in pH readings in the two media was 0.04 pH units. In summary, we have demonstrated that these optical sensors agree well with standard electrochemical probes. The accuracy of the optical probes demonstrates their ability to detect potential parameter drift that could have significant impact on growth, production kinetics, and protein product quality. We have also shown that an increase in osmolality that could result from controlling pH or operating the reactor in fed-batch mode has an insignificant impact on the functionality of the pH patches.     

Single-stranded DNAzyme-based Pb2+ fluorescent sensor that can work well over a wide temperature range

DNAzymes have become an excellent choice for sensing applications. Based on DNAzymes, three generations of Pb(2+) fluorescent sensors have been reported. In these sensors, two oligonucleotide strands (substrate strand and enzyme strand) were used, which not only increased the complexity of the detection system, but also brought some difficulties for the use of the sensors at elevated temperatures. To overcome this problem, a single-stranded DNAzyme-based Pb(2+) fluorescent sensor was designed by combining the substrate sequence and the enzyme sequence into one oligonucleotide strand. The intramolecular duplex structure of this single-stranded DNAzyme kept the fluorophore and the quencher, labeled at its two ends, in close proximity; thus the background fluorescence was significantly suppressed. Using this fluorescent sensor, Pb(2+) quantitation can be achieved with high sensitivity and high selectivity. In addition, the extraordinary stability of the intramolecular duplex structure could assure a low background fluorescence at high temperature, even if the number of complementary base pairs between the substrate sequence and the enzyme sequence was reduced, allowing the sensor to work well over a wide temperature range. Similar performances of the fluorescent sensor at 4, 25 and 37°C suggested that this sensor has a good ability to resist temperature fluctuations.     

Interferometric optical fiber sensor for optoacoustic endomicroscopy

Optical fiber sensors can offer robust and miniaturized detection of wideband ultrasound, yielding high sensitivity and immunity to electromagnetic interference. However, the lack of cost-effective manufacturing methods prevents the disseminated use of these sensors in biomedical applications. In this study, we developed and optimized a simple method to create optical cavities with high-quality mirrors for acoustic sensing based on micro-manipulation of UV-curable optical adhesives and electroless chemical silver deposition. This approach enables the manufacturing of ultrasound sensors based on Fabry-Pérot interferometers on optical fiber tips with minimal production costs. Characterization and high-resolution optoacoustic imaging experiments show that the manufacturing process yielded a fiber sensor with a small NEP () over a broad detection bandwidth (25 MHz), generally outperforming conventional piezoelectric based transducers. We discuss how the new manufacturing process leads to a high-performance acoustic detector that, due to low cost, can be used as a disposable sensor.