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Extracellular acidification is a basic indicator for alterations in two vital metabolic pathways: glycolysis and cellular respiration. Measuring these alterations by monitoring extracellular acidification using cell-based biosensors such as LAPS plays an important role in studying these pathways whose disorders are associated with numerous diseases including cancer. However, the surface of the biosensors must be specially tailored to ensure high cell compatibility so that cells can represent more in vivo-like behavior, which is critical to gain more realistic in vitro results from the analyses, e.g., drug discovery experiments. In this work, O2 plasma patterning on the LAPS surface is studied to enhance surface features of the sensor chip, e.g., wettability and biofunctionality. The surface treated with O2 plasma for 30 s exhibits enhanced cytocompatibility for adherent CHO–K1 cells, which promotes cell spreading and proliferation. The plasma-modified LAPS chip is then integrated into a microfluidic system, which provides two identical channels to facilitate differential measurements of the extracellular acidification of CHO–K1 cells. To the best of our knowledge, it is the first time that extracellular acidification within microfluidic channels is quantitatively visualized as differential (bio-)chemical images.
A light-addressable potentiometric sensor (LAPS) is a field-effect-based (bio-) chemical sensor, in which a desired sensing area on the sensor surface can be defined by illumination. Light addressability can be used to visualize the concentration and spatial distribution of the target molecules, e.g., H+ ions. This unique feature has great potential for the label-free imaging of the metabolic activity of living organisms. The cultivation of those organisms needs specially tailored surface properties of the sensor. O2 plasma treatment is an attractive and promising tool for rapid surface engineering. However, the potential impacts of the technique are carefully investigated for the sensors that suffer from plasma-induced damage. Herein, a LAPS with a Ta2O5 pH-sensitive surface is successfully patterned by plasma treatment, and its effects are investigated by contact angle and scanning LAPS measurements. The plasma duration of 30 s (30 W) is found to be the threshold value, where excessive wettability begins. Furthermore, this treatment approach causes moderate plasma-induced damage, which can be reduced by thermal annealing (10 min at 300 °C). These findings provide a useful guideline to support future studies, where the LAPS surface is desired to be more hydrophilic by O2 plasma treatment.
As one class of molecular imprinted polymers (MIPs), surface imprinted polymer (SIP)-based biosensors show great potential in direct whole-bacteria detection. Micro-contact imprinting, that involves stamping the template bacteria immobilized on a substrate into a pre-polymerized polymer matrix, is the most straightforward and prominent method to obtain SIP-based biosensors. However, the major drawbacks of the method arise from the requirement for fresh template bacteria and often non-reproducible bacteria distribution on the stamp substrate. Herein, we developed a positive master stamp containing photolithographic mimics of the template bacteria (E. coli) enabling reproducible fabrication of biomimetic SIP-based biosensors without the need for the “real” bacteria cells. By using atomic force and scanning electron microscopy imaging techniques, respectively, the E. coli-capturing ability of the SIP samples was tested, and compared with non-imprinted polymer (NIP)-based samples and control SIP samples, in which the cavity geometry does not match with E. coli cells. It was revealed that the presence of the biomimetic E. coli imprints with a specifically designed geometry increases the sensor E. coli-capturing ability by an “imprinting factor” of about 3. These findings show the importance of geometry-guided physical recognition in bacterial detection using SIP-based biosensors. In addition, this imprinting strategy was employed to interdigitated electrodes and QCM (quartz crystal microbalance) chips. E. coli detection performance of the sensors was demonstrated with electrochemical impedance spectroscopy (EIS) and QCM measurements with dissipation monitoring technique (QCM-D).
Recognition of subjects with mild cognitive impairment (MCI) by the use of retinal arterial vessels.
(2019)
The ClearPET™ project is proposed by working groups of the Crystal Clear Collaboration (CCC) to develop a 2nd generation high performance small animal positron emission tomograph (PET). High sensitivity and high spatial resolution is foreseen for the ClearPET™ camera by using a phoswich arrangement combining mixed lutetium yttrium aluminum perovskite (LuYAP:Ce) and lutetium oxyorthosilicate (LSO) scintillating crystals. Design optimizations for the first photomultiplier tube (PMT) based ClearPET camera are done with a Monte-Carlo simulation package implemented on GEANT3 (CERN, Geneva, Switzerland). A dual-head prototype has been built to test the frontend electronics and was used to validate the implementation of the GEANT3 simulation tool. Multiple simulations were performed following the experimental protocols to measure the intrinsic resolution and the sensitivity profile in axial and radial direction. Including a mean energy resolution of about 27.0% the simulated intrinsic resolution is about (1.41±0.11)mm compared to the measured of (1.48±0.06)mm. The simulated sensitivity profiles show a mean square deviation of 12.6% in axial direction and 3.6% in radial direction. Satisfactorily these results are representative for all designs and confirm the scanner geometry.
Beyond ClearPET: Next Aims
(2008)
The CRYSTAL CLEAR collaboration, in short CCC, is a consortium of 12 academic institutions, mainly from Europe, joining efforts in the area of developing instrumentation for nuclear medicine and medical imaging. In the framework of the CCC a high performance small animal PET system, called ClearPET, was developed by using new technologies in electronics and crystals in a phoswich arrangement combining two types of lutetium- based scintillator materials: LSO:Ce and LuYAP:Ce. Our next aim will be the development of hybrid image systems. Hybrid MR-PET imaging has many unique advantages for brain research. This has sparked a new research line within CCC for the development of novel MR-PET compatible technologies. MRI is not as sensitive as PET but PET has poorer spatial resolution than MRI. Two major advantages of PET are sensitivity and its ability to acquire metabolic information. To assess these innovations, the development of a 9.4T hybrid animal MR-PET scanner is proposed based on an existing 9.4T MR scanner that will be adapted to enable simultaneous acquisition of MR and PET data using cutting- edge technology for both MR and PET.
The ClearPET™ project: Development of a 2nd generation high-performance small animal PET scanner
(2005)
Second generation high-performance PET scanners, called ClearPET™1, have been developed by working groups of the Crystal Clear Collaboration (CCC). High sensitivity and high spatial resolution for the ClearPET camera is achieved by using a phoswich arrangement combining two different types of lutetium-based scintillator materials: LSO from CTI and LuYAP:Ce from the CCC (ISTC project). In a first ClearPET prototype, phoswich arrangements of 8×8 crystals of 2×2×10 mm3 are coupled to multi-channel photomultiplier tubes (Hamamatsu R7600). A unit of four PMTs arranged in-line represents one of 20 sectors of the ring design. The opening diameter of the ring is 120 mm, the axial detector length is 110 mm.The PMT pulses are digitized by free-running ADCs and digital data processing determines the gamma energy, the phoswich layer and even the exact pulse starting time, which is subsequently used for coincidence detection. The gantry allows rotation of the detector modules around the field of view.
Preliminary data shows a correct identification of the crystal layer about (98±1)%. Typically the energy resolution is (23.3±0.5)% for the luyap layer and (15.4±0.4)% for the lso layer. early studies showed the timing resolution of 2 ns FWHM and 4.8 ns FWTM. the intrinsic spatial resolution ranges from 1.37 mm to 1.61 mm full-width of half-maximum (FWHM) with a mean of 1.48 mm FWHM. further improvements in image and energy resolution are expected when the system geometry is fully modeled.
A 2nd generation high performance small animal PET scanner, called ClearPET™, has been designed and a first prototype is built by working groups of the Crystal Clear Collaboration (CCC). In order to achieve high sensitivity and maintain good uniform spatial resolution over the field of view in high resolution PET systems, it is necessary to extract the depth of interaction (DOI) information and correct for spatial degradation. The design of the first ClearPET™ Demonstrator based on the use of the multi-anode photomultiplier tube (Hamamatsu R7600-M64) and a LSO/LuYAP phoswich matrix. The two crystal layers of 8*8 crystals (2*2*10 mm3) are stacked on each other and mounted without light guide as one to one on the PMT. A unit of four PMTs arranged in-line represents one of 20 sectors of the ring design. The opening diameter of the crystal ring is 137 mm, the axial detector length is 110 mm. The PMT pulses are digitized by free-running ADCs and digital data processing determines the gamma energy, the phoswich layer and even the pulse arrival time. Single gamma interactions are recorded and coincidences are found by software. The gantry allows rotation of the detector modules around the field of view. The measurements have been done using the first LSO/LuYAP detector cassettes.
Differential multiplicities of forward produced hadrons in deep inelastic muon scattering on nuclear targets have been compared with those from deuterium. The ratios are observed to increase towards unity as the virtual photon energy increases with no significant dependence on the other muon kinematic variables. The hadron transverse momentum distribution is observed to be broadened in nuclear targets. The dependence on the remaining hadron variables is investigated and the results are discussed in the framework of intranuclear interaction models and in the context of the EMC effect.
Measurements are presented of the inclusive distributions of the J/Ψ meson produced by muons of energy 200 GeV from an ammonia target. The gluon distribution of the nucleon has been derived from the data in the range 0.04<x<0.36 using a technique based on the colour singlet model. An arbitrary normalisation factor is required to obtain a reasonable integral of the gluon distribution. Some comments are made on the use of J/Ψ productionby virtual photons to extract the gluon distribution at HERA.
Results are presented on the ratios of the nucleon structure function in copper to deuterium from two separate experiments. The data confirm that the nucleon structure function,F 2, is different for bound nucleons than for the quasi-free ones in the deuteron. The redistribution in the fraction of the nucleon's momentum carried by quarks is investigated and it is found that the data are compatible with no integral loss of quark momenta due to nuclear effects.
The spin asymmetry in deep inelastic scattering of longitudinally polarised muons by longitudinally polarised protons has been measured over a large x range (0.01<x<0.7). The spin-dependent structure function g1(x) for the proton has been determined and its integral over x found to be 0.114±0.012±0.026, in disagreement with the Ellis-Jaffe sum rule. Assuming the validity of the Bjorken sum rule, this result implies a significant negative value for the integral of g1 for the neutron. These values for the integrals of g1 lead to the conclusion that the total quark spin constitutes a rather small fraction of the spin of the nucleon.
The spin asymmetry in deep inelastic scattering of longitudinally polarised muons by longitudinally polarised protons has been measured in the range 0.01<×<0.7. The spin dependent structure function g1(x) for the proton has been determined and, combining the data with earlier SLAC measurements, its integral over x found to be 0.126±0.010(stat.)±0.015(syst.), in disagreement with the Ellis-Jaffe sum rule. Assuming the validity of the Biorken sum rule, this result implies a significant negative value for the integral of g1 for the neutron. These integrals lead to the conclusion, in the naïve quark parton model, that the total quark spin constitutes a rather small fraction of the spin of the nucleon. Results are also presented on the asymmetries in inclusive hadron production which are consistent with the above picture.
This chapter shows that nanomaterials obtained by high-temperature carbonization of inexpensive plant raw material such as rice husk, grape seeds, and walnut shells can serve as a basis for the production of highly efficient microbial drugs, biodestructors, biosorbents, and biocatalysts, which are promising for the remediation of the ecosystem contaminated with heavy and radioactive metals, oil and oil products. A strong interest in engineering zymology is dictated by the necessity to address the issues of monitoring enzymatic processes, treatment, and diagnosis of a number of common human diseases, environmental pollution, quality control of pharmaceuticals and food. Nanomaterials obtained by high-temperature carbonization of cheap plant raw material such as-rice husks, grape seeds and walnut shells, can serve as a basis for creating of highly effective microbial preparations-biodestructors, biosorbents and biocatalysts, which are promising for the use of contaminated ecosystems, and for restoration of human intestine microecology.
The most of conventional methods of air purification use the power of a fan to draw in air and pass it through a filter. The problem of bacterial contamination of inner parts of such a type of air conditioners in some cases draws attention towards alternative air-cleaning systems. Some manufacturers offer to use the ozone's bactericidal and deodorizing effects, but the wide spreading of such systems is restricted by the fact that toxic effects of ozone in respect of human beings are well known. In 2000 Sharp Inc. introduced "Plasma Cluster Ions (PCI)" air purification technology, which uses plasma discharge to generate cluster ions (I 0-14 ). This technology has been developed for those customers that are conscious about health and hygiene. In our experiments, we focused on some principal aspects of plasma-generated ions application - time-dependency and irreversibility of bactericidal action, spatial and kinetic characteristics of emitted cluster particles, their chemical targets in the microbial cells.
Bacterial lipopolysaccharides (endotoxins) show strong biological effects at very low concentrations in human beings and many animals when entering the blood stream. These include affecting structure and function of organs and cells, changing metabolic functions, raising body temperature, triggering the coagulation cascade, modifying hemodynamics and causing septic shock. Because of this toxicity, the removal of even minute amounts is essential for safe parenteral administration of drugs and also for septic shock patients' care. The absence of a general method for endotoxin removal from liquid interfaces urgently requires finding new methods and materials to overcome this gap. Nanostructured carbonized plant parts is a promising material that showed good adsorption properties due to its vast pore network and high surface area. The aim of this study was comparative measurement of endotoxin- and blood proteins-related adsorption rate and adsorption capacity for different carboneous materials produced at different temperatures and under different surface modifications. As a main surface modificator, positively cbarged polymer, polyethileneimine (PEl) was used. Activated carbon materials showed good adsorption properties for LPS and some proteins used in the experiments. During the batch experiments, several techniques (dust removal, autoclaving) were used and optimized for improving the material's adsorption behavior. Also, with the results obtained it was possible to differentiate the materials according to their adsorption capacity and kinetic characteristics. Modification of the surface apparently has not affected hemoglobin binding to the adsorbent's surface. Obtained adsorption isotherms can be used as a powerful tool for designing of future column-based setups for blood purification from LPS, which is especially important for septic shock treatment.
The deformation and damage laws of non-homogeneous irregular structural planes in rocks are the basis for studying the stability of rock engineering. To investigate the damage characteristics of rock containing non-parallel fissures, uniaxial compression tests and numerical simulations were conducted on sandstone specimens containing three non-parallel fissures inclined at 0°, 45° and 90° in this study. The characteristics of crack initiation and crack evolution of fissures with different inclinations were analyzed. A constitutive model for the discontinuous fractures of fissured sandstone was proposed. The results show that the fracture behaviors of fissured sandstone specimens are discontinuous. The stress–strain curves are non-smooth and can be divided into nonlinear crack closure stage, linear elastic stage, plastic stage and brittle failure stage, of which the plastic stage contains discontinuous stress drops. During the uniaxial compression test, the middle or ends of 0° fissures were the first to crack compared to 45° and 90° fissures. The end with small distance between 0° and 45° fissures cracked first, and the end with large distance cracked later. After the final failure, 0° fissures in all specimens were fractured, while 45° and 90° fissures were not necessarily fractured. Numerical simulation results show that the concentration of compressive stress at the tips of 0°, 45° and 90° fissures, as well as the concentration of tensile stress on both sides, decreased with the increase of the inclination angle. A constitutive model for the discontinuous fractures of fissured sandstone specimens was derived by combining the logistic model and damage mechanic theory. This model can well describe the discontinuous drops of stress and agrees well with the whole processes of the stress–strain curves of the fissured sandstone specimens.
Light-addressable potentiometric sensors for quantitative spatial imaging of chemical species
(2017)
A light-addressable potentiometric sensor (LAPS) is a semiconductor-based chemical sensor, in which a measurement site on the sensing surface is defined by illumination. This light addressability can be applied to visualize the spatial distribution of pH or the concentration of a specific chemical species, with potential applications in the fields of chemistry, materials science, biology, and medicine. In this review, the features of this chemical imaging sensor technology are compared with those of other technologies. Instrumentation, principles of operation, and various measurement modes of chemical imaging sensor systems are described. The review discusses and summarizes state-of-the-art technologies, especially with regard to the spatial resolution and measurement speed; for example, a high spatial resolution in a submicron range and a readout speed in the range of several tens of thousands of pixels per second have been achieved with the LAPS. The possibility of combining this technology with microfluidic devices and other potential future developments are discussed.
The light-addressable potentiometric sensor (LAPS) is an electrochemical sensor with a field-effect structure to detect the variation of the Nernst potential at its sensor surface, the measured area on which is defined by illumination. Thanks to this light-addressability, the LAPS can be applied to chemical imaging sensor systems, which can visualize the two-dimensional distribution of a particular target ion on the sensor surface. Chemical imaging sensor systems are expected to be useful for analysis of reaction and diffusion in various electrochemical and biological samples. Recent developments of LAPS-based chemical imaging sensor systems, in terms of the spatial resolution, measurement speed, image quality, miniaturization and integration with microfluidic devices, are summarized and discussed.
The artificial olfactory image was proposed by Lundström et al. in 1991 as a new strategy for an electronic nose system which generated a two-dimensional mapping to be interpreted as a fingerprint of the detected gas species. The potential distribution generated by the catalytic metals integrated into a semiconductor field-effect structure was read as a photocurrent signal generated by scanning light pulses. The impact of the proposed technology spread beyond gas sensing, inspiring the development of various imaging modalities based on the light addressing of field-effect structures to obtain spatial maps of pH distribution, ions, molecules, and impedance, and these modalities have been applied in both biological and non-biological systems. These light-addressing technologies have been further developed to realize the position control of a faradaic current on the electrode surface for localized electrochemical reactions and amperometric measurements, as well as the actuation of liquids in microfluidic devices.
The light-addressable potentiometric sensor (LAPS) and scanning photo-induced impedance microscopy (SPIM) are two closely related methods to visualise the distributions of chemical species and impedance, respectively, at the interface between the sensing surface and the sample solution. They both have the same field-effect structure based on a semiconductor, which allows spatially resolved and label-free measurement of chemical species and impedance in the form of a photocurrent signal generated by a scanning light beam. In this article, the principles and various operation modes of LAPS and SPIM, functionalisation of the sensing surface for measuring various species, LAPS-based chemical imaging and high-resolution sensors based on silicon-on-sapphire substrates are described and discussed, focusing on their technical details and prospective applications.
An application of a scanning light-addressable potentiometric sensor for label-free DNA detection
(2013)
A multi-spot light-addressable potentiometric sensor (LAPS), which belongs to the family of semiconductor field-effect devices, was applied for label-free detection of double-stranded deoxyribonucleic acid (dsDNA) molecules by their intrinsic molecular charge. To reduce the distance between the DNA charge and sensor surface and thus, to enhance the electrostatic coupling between the dsDNA molecules and the LAPS, the negatively charged dsDNA molecules were electrostatically adsorbed onto the gate surface of the LAPS covered with a positively charged weak polyelectrolyte layer of PAH (poly(allylamine hydrochloride)). The surface potential changes in each spot of the LAPS, induced by the layer-by-layer adsorption of a PAH/dsDNA bilayer, were recorded by means of photocurrent-voltage and constant-photocurrent measurements. In addition, the surface morphology of the gate surface before and after consecutive electrostatic adsorption of PAH and dsDNA layers was studied by atomic force microscopy measurements. Moreover, fluorescence microscopy was used to verify the successful adsorption of dsDNA molecules onto the PAH-modified LAPS surface. A high sensor signal of 25 mV was registered after adsorption of 10 nM dsDNA molecules. The lower detection limit is down to 0.1 nM dsDNA. The obtained results demonstrate that the PAH-modified LAPS device provides a convenient and rapid platform for the direct label-free electrical detection of in-solution hybridized dsDNA molecules.