@article{MuschallikMolinnusBongaertsetal.2017, author = {Muschallik, Lukas and Molinnus, Denise and Bongaerts, Johannes and Pohl, Martina and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Siegert, Petra and Selmer, Thorsten}, title = {(R,R)-Butane-2,3-diol Dehydrogenase from Bacillus clausii DSM 8716T: Cloning and Expression of the bdhA-Gene, and Initial Characterization of Enzyme}, series = {Journal of Biotechnology}, volume = {258}, journal = {Journal of Biotechnology}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0168-1656}, doi = {10.1016/j.jbiotec.2017.07.020}, pages = {41 -- 50}, year = {2017}, abstract = {The gene encoding a putative (R,R)-butane-2,3-diol dehydrogenase (bdhA) from Bacillus clausii DSM 8716T was isolated, sequenced and expressed in Escherichia coli. The amino acid sequence of the encoded protein is only distantly related to previously studied enzymes (identity 33-43\%) and exhibited some uncharted peculiarities. An N-terminally StrepII-tagged enzyme variant was purified and initially characterized. The isolated enzyme catalyzed the (R)-specific oxidation of (R,R)- and meso-butane-2,3-diol to (R)- and (S)-acetoin with specific activities of 12 U/mg and 23 U/mg, respectively. Likewise, racemic acetoin was reduced with a specific activity of up to 115 U/mg yielding a mixture of (R,R)- and meso-butane-2,3-diol, while the enzyme reduced butane-2,3-dione (Vmax 74 U/mg) solely to (R,R)-butane-2,3-diol via (R)-acetoin. For these reactions only activity with the co-substrates NADH/NAD+ was observed. The enzyme accepted a selection of vicinal diketones, α-hydroxy ketones and vicinal diols as alternative substrates. Although the physiological function of the enzyme in B. clausii remains elusive, the data presented herein clearly demonstrates that the encoded enzyme is a genuine (R,R)-butane-2,3-diol dehydrogenase with potential for applications in biocatalysis and sensor development.}, language = {en} } @article{YoshinobuMiyamotoWerneretal.2017, author = {Yoshinobu, Tatsuo and Miyamoto, Ko-ichiro and Werner, Frederik and Poghossian, Arshak and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Light-addressable potentiometric sensors for quantitative spatial imaging of chemical species}, series = {Annual Review of Analytical Chemistry}, volume = {10}, journal = {Annual Review of Analytical Chemistry}, publisher = {Annual Reviews}, address = {Palo Alto, Calif.}, issn = {1936-1327}, doi = {10.1146/annurev-anchem-061516-045158}, pages = {225 -- 246}, year = {2017}, abstract = {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.}, language = {en} } @article{PoghossianWernerBuniatyanetal.2017, author = {Poghossian, Arshak and Werner, Frederik and Buniatyan, V. V. and Wagner, Torsten and Miamoto, K. and Yoshinobu, T. and Sch{\"o}ning, Michael Josef}, title = {Towards addressability of light-addressable potentiometric sensors: Shunting effect of non-illuminated region and cross-talk}, series = {Sensor and Actuators B: Chemical}, journal = {Sensor and Actuators B: Chemical}, number = {244}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2017.01.047}, pages = {1071 -- 1079}, year = {2017}, abstract = {The LAPS (light-addressable potentiometric sensor) platform is one of the most attractive approaches for chemical and biological sensing with many applications ranging from pH and ion/analyte concentration measurements up to cell metabolism detection and chemical imaging. However, although it is generally accepted that LAPS measurements are spatially resolved, the light-addressability feature of LAPS devices has not been discussed in detail so far. In this work, an extended electrical equivalent-circuit model of the LAPS has been presented, which takes into account possible cross-talk effects due to the capacitive coupling of the non-illuminated region. A shunting effect of the non-illuminated area on the measured photocurrent and addressability of LAPS devices has been studied. It has been shown, that the measured photocurrent will be determined not only by the local interfacial potential in the illuminated region but also by possible interfacial potential changes in the non-illuminated region, yielding cross-talk effects. These findings were supported by the experimental investigations of a penicillin-sensitive multi-spot LAPS and a metal-insulator-semiconductor LAPS as model systems.}, language = {en} } @article{DollWagnerWagneretal.2016, author = {Doll, Theodor and Wagner, Torsten and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Engineering of functional interfaces / Theodor Doll ; Torsten Wagner ; Patrick Wagner ; Michael J. Sch{\"o}ning (eds.)}, series = {Physica status solidi (a)}, volume = {213}, journal = {Physica status solidi (a)}, number = {6}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201670641}, pages = {1393 -- 1394}, year = {2016}, language = {en} } @article{WagnerVornholtWerneretal.2016, author = {Wagner, Torsten and Vornholt, Wolfgang and Werner, Frederik and Yoshinobu, Tatsuo and Miyamoto, Ko-Ichiro and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Light-addressable potentiometric sensor (LAPS) combined with magnetic beads for pharmaceutical screening}, series = {Physics in medicine}, volume = {2016}, journal = {Physics in medicine}, number = {1}, issn = {2352-4510}, doi = {10.1016/j.phmed.2016.03.001}, pages = {2 -- 7}, year = {2016}, abstract = {The light-addressable potentiometric sensor (LAPS) has the unique feature to address different regions of a sensor surface without the need of complex structures. Measurements at different locations on the sensor surface can be performed in a common analyte solution, which distinctly simplifies the fluidic set-up. However, the measurement in a single analyte chamber prevents the application of different drugs or different concentrations of a drug to each measurement spot at the same time as in the case of multi-reservoir-based set-ups. In this work, the authors designed a LAPS-based set-up for cell culture screening that utilises magnetic beads loaded with the endotoxin (lipopolysaccharides, LPS), to generate a spatially distributed gradient of analyte concentration. Different external magnetic fields can be adjusted to move the magnetic beads loaded with a specific drug within the measurement cell. By recording the metabolic activities of a cell layer cultured on top of the LAPS surface, this work shows the possibility to apply different concentrations of a sample along the LAPS measurement spots within a common analyte solution.}, language = {en} } @article{MiyamotoSatoAbeetal.2016, author = {Miyamoto, Ko-Ichiro and Sato, Takuya and Abe, Minami and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Light-addressable potentiometric sensor as a sensing element in plug-based microfluidic devices}, series = {Micromachines}, volume = {7}, journal = {Micromachines}, number = {7}, publisher = {MDPI}, address = {Basel}, issn = {2072-666X}, doi = {10.3390/mi7070111}, pages = {111}, year = {2016}, abstract = {A plug-based microfluidic system based on the principle of the light-addressable potentiometric sensor (LAPS) is proposed. The LAPS is a semiconductor-based chemical sensor, which has a free addressability of the measurement point on the sensing surface. By combining a microfluidic device and LAPS, ion sensing can be performed anywhere inside the microfluidic channel. In this study, the sample solution to be measured was introduced into the channel in a form of a plug with a volume in the range of microliters. Taking advantage of the light-addressability, the position of the plug could be monitored and pneumatically controlled. With the developed system, the pH value of a plug with a volume down to 400 nL could be measured. As an example of plug-based operation, two plugs were merged in the channel, and the pH change was detected by differential measurement.}, language = {en} } @article{BreuerRaueStrobeletal.2016, author = {Breuer, Lars and Raue, Markus and Strobel, M. and Mang, Thomas and Sch{\"o}ning, Michael Josef and Thoelen, R. and Wagner, Torsten}, title = {Hydrogels with incorporated graphene oxide as light-addressable actuator materials for cell culture environments in lab-on-chip systems}, series = {Physica status solidi (a)}, volume = {213}, journal = {Physica status solidi (a)}, number = {6}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6300}, doi = {10.1002/pssa.201533056}, pages = {1520 -- 1525}, year = {2016}, abstract = {Abstractauthoren Graphene oxide (GO) nanoparticles were incorporated in temperature-sensitive Poly(N-isopropylacrylamide) (PNIPAAm) hydrogels. The nanoparticles increase the light absorption and convert light energy into heat efficiently. Thus, the hydrogels with GO can be stimulated spatially resolved by illumination as it was demonstrated by IR thermography. The temporal progression of the temperature maximum was detected for different concentrations of GO within the polymer network. Furthermore, the compatibility of PNIPAAm hydrogels with GO and cell cultures was investigated. For this purpose, culture medium was incubated with hydrogels containing GO and the viability and morphology of chinese hamster ovary (CHO) cells was examined after several days of culturing in presence of this medium.}, language = {en} } @article{DantismTakenagaWagneretal.2016, author = {Dantism, Shahriar and Takenaga, Shoko and Wagner, Patrick and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Determination of the extracellular acidification of Escherichia coli K12 with a multi-​chamber-​based LAPS system}, series = {Physica status solidi (a)}, volume = {213}, journal = {Physica status solidi (a)}, number = {6}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6300}, doi = {10.1002/pssa.201533043}, pages = {1479 -- 1485}, year = {2016}, abstract = {On-line monitoring of the metabolic activity of microorganisms involved in intermediate stages of biogas production plays an important role to avoid undesirable "down times" during the biogas production. In order to control this process, an on-chip differential measuring system based on the light-addressable potentiometric sensor (LAPS) principle combined with a 3D-printed multi-chamber structure has been realized. As a test microorganism, Escherichia coli K12 (E. coli K12) were used for cell-based measurements. Multi-chamber structures were developed to determine the metabolic activity of E. coli K12 in suspension for a different number of cells, responding to the addition of a constant or variable amount of glucose concentrations, enabling differential and simultaneous measurements.}, language = {en} } @article{MiyamotoYuIsodaetal.2016, author = {Miyamoto, Ko-ichiro and Yu, Bing and Isoda, Hiroko and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Visualization of the recovery process of defects in a cultured cell layer by chemical imaging sensor}, series = {Sensors and Actuators B: Chemical}, volume = {236}, journal = {Sensors and Actuators B: Chemical}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2016.04.018}, pages = {965 -- 969}, year = {2016}, abstract = {The chemical imaging sensor is a field-effect sensor which is able to visualize both the distribution of ions (in LAPS mode) and the distribution of impedance (in SPIM mode) in the sample. In this study, a novel cell assay is proposed, in which the chemical imaging sensor operated in SPIM mode is applied to monitor the recovery of defects in a cell layer brought into proximity of the sensing surface. A reduced impedance at a defect formed artificially in a cell layer was successfully visualized in a photocurrent image. The cell layer was cultured over two weeks, during which the temporal change of the photocurrent distribution corresponding to the recovery of the defect was observed.}, language = {de} } @article{MiyamatoSakakitaWagneretal.2015, author = {Miyamato, Ko-ichiro and Sakakita, Sakura and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Application of chemical imaging sensor to in-situ pH imaging in the vicinity of a corroding metal surface}, series = {Electrochimica Acta}, volume = {183}, journal = {Electrochimica Acta}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0013-4686}, doi = {10.1016/j.electacta.2015.07.184}, pages = {137 -- 142}, year = {2015}, abstract = {The chemical imaging sensor was applied to in-situ pH imaging of the solution in the vicinity of a corroding surface of stainless steel under potentiostatic polarization. A test piece of polished stainless steel was placed on the sensing surface leaving a narrow gap filled with artificial seawater and the stainless steel was corroded under polarization. The pH images obtained during polarization showed correspondence between the region of lower pH and the site of corrosion. It was also found that the pH value in the gap became as low as 2 by polarization, which triggered corrosion.}, language = {en} }