| 1. |
- Spegel, Christer, et al.
(author)
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On-chip determination of dopamine exocytosis using mercaptopropionic acid modified microelectrodes
- 2007
-
In: Electroanalysis. - : Wiley. - 1040-0397 .- 1521-4109. ; 19:2-3, s. 263-271
-
Journal article (peer-reviewed)abstract
- Gold and platinum, which often are used for thin film metallization, are not suitable for the measurement of dopamine (DA), since the oxidation product of DA forms a non-conducting polymer on the electrode surface. In this work several thiols were screened for their ability to prevent this polymerization. It was found that mercaptopropionic acid (MPA) decreased the rate of DA polymerization. MPA, possessing a weak acidic functionality, had the greatest effect on the DA electrochemistry by decreasing electrode passivation, as well as improving reversibility and sensitivity. Modifications of microchip electrodes with MPA did not only improve DA electrochemistry but also significantly increased the storage stability of the transducers. The microchips were ultimately used to detect K+ stimulated quantal release of DA from PC12 cells.
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| 2. |
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| 3. |
- Badea, M, et al.
(author)
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A flow immunoassay for alkylphenol ethoxylate surfactants and their metabolites - questions associated with cross-reactivity, matrix effects, and validation by chromatographic techniques
- 2003
-
In: Analyst. - : Royal Society of Chemistry (RSC). - 1364-5528. ; 128:7, s. 849-856
-
Journal article (peer-reviewed)abstract
- This paper describes the application and evaluation of a competitive enzyme flow injection immunoassay (EFIIA) for screening of alkylphenol ethoxylate (APEO) surfactants in different water samples based on a generic immunoassay system previously developed ( see E. Burestedt, C Nistor, U. Schagerlof and J. Emneus, Anal. Chem., 2000, 72, 4171 - 4177). The detection limits for octylphenol ethoxylates (OPEOs), nonylphenol ethoxylates (NPEOs), and nonylphenol (NP) were 0.5 mug l(-1), between 2 and 3 mug l(-1), and 50 mug l(-1), respectively, with a sample throughput of 6 h(-1) (i.e., for triplicate analysis of each sample). Different OPEOs and NPEOs were highly cross-reactive within the assay, with sensitivities in the same order of magnitude for all the ethoxylates tested, thus the result obtained by the EFIIA method could be used as an "alkylphenol ethoxylate index". No or minor matrix effects with recoveries between 70 - 120% for the reference analyte NPEO10 in tap, and surface water, and acceptable for rainwater, were observed. Influent and effluent surfactant containing wastewater samples were analysed by EFIIA, LC-MS, LC-Fluoresence (LC-FL), and a commercial microplate ELISA. High recoveries for different concentrations of APEO(10) spiked into a 200 times diluted raw influent and effluent wastewater were achieved with the EFIIA method, however, the found APEO content of the same diluted wastewater samples, before spiking, could not be correlated directly to the chromatographic result by any of the immunoassays, and the possible reasons for this are discussed. The same trend of decreasing APEO content from influent to effluent wastewater could, however, be followed for all methods employed.
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| 4. |
- Bjarnason, Bjarni, et al.
(author)
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Enzyme flow immunoassay using a Protein G column for the screening of triazine herbicides in surface and waste water
- 2001
-
In: Analytica Chimica Acta. - 0003-2670. ; 426:2, s. 197-207
-
Journal article (peer-reviewed)abstract
- A method for screening of triazine herbicides in surface and waste water is presented. The method is based on an enzyme flow immunoassay (EFIA) for the detection of the free fraction of a horse radish peroxidase (HRP)-labelled antigen (tracer). This was accomplished by trapping the bound tracer fraction in a Protein G column, allowing the residual free tracer fraction to pass and be detected spectrophotometrically after incubation with an enzyme substrate. As compared with detecting the bound tracer fraction this reduces the regeneration requirements of the Protein G column used for capturing the bound fraction and, therefore, reduces assay time. A polyclonal antibody directed against simazine showed no reactivity towards tracers that were thiopropionic acid derivatives of atrazine, simazine and terbutylazine. It had good sensitivity towards tracers using derivatives of 2-chloro-4,6-(alkylamino)-s-triazines such as atrazine and simazine. The highest sensitivity was obtained with an Et/Cl/N-C5-HRP tracer because this tracer could be used in combination with the lowest concentration of antibody. The detection limit was 0.1 μgl-1 with a linear range between 0.1 and 10 μgl-1 and an assay throughput of 12 h-1. Natural water samples from various locations in Russia were analysed for triazines and the results were compared with a previously developed fluorescein flow immunoassay for triazines. The results were further verified by supported liquid membrane (SLM) extraction combined with HPLC. The results show that the two immunoassays behave differently and that the sample matrix influences their performance, however, no false negative results were obtained. The possible reasons for the different results between the two immunoassays are discussed. (C) 2001 Elsevier Science B.V.
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| 5. |
- Bunea, Ada-Ioana, et al.
(author)
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Micropatterned Carbon-on-Quartz Electrode Chips for Photocurrent Generation from Thylakoid Membranes
- 2018
-
In: ACS Applied Energy Materials. - : American Chemical Society (ACS). - 2574-0962. ; 1:7, s. 3313-3322
-
Journal article (peer-reviewed)abstract
- Harvesting the energy generated by photosynthetic organisms through light-dependent reactions is a significant step toward a sustainable future energy supply. Thylakoid membranes are the site of photosynthesis, and thus particularly suited for developing photo-bioelectrochemical cells. Novel electrode materials and geometries could potentially improve the efficiency of energy harvesting using thylakoid membranes. For commercial applications, electrodes with large surface areas are needed. Photolithographic patterning of a photoresist, followed by pyrolysis, is a flexible and fast approach for the fabrication of carbon electrodes with tailored properties. In this work, electrode chips consisting of patterned carbon supported on quartz were designed and fabricated. The patterned electrode area is 1 cm2, and the measurement chamber footprint is 0.5 cm2, 1 order of magnitude larger than previously tested electrodes for thylakoid membrane immobilization. The use of a transparent substrate allows back-side illumination, protecting the bioelectrochemical system from the environment and vice versa. Two different mediators, monomeric ([Ru(NH3)6]3+) and polymeric ([Os(2,2′-bipyridine)2-poly(N-vinylimidazole)10Cl]+/2+), are used for evaluating photocurrent generation from thylakoid membranes with different electrode geometries. Current densities up to 71 μA cm–2 are measured upon illumination through the transparent electrode chip with solar simulated irradiance (1000 W m–2).
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| 6. |
- Cunha, André B., et al.
(author)
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Development of a Smart Wireless Multisensor Platform for an Optogenetic Brain Implant
- 2024
-
In: Sensors. - 1424-8220. ; 24:2
-
Journal article (peer-reviewed)abstract
- Implantable cell replacement therapies promise to completely restore the function of neural structures, possibly changing how we currently perceive the onset of neurodegenerative diseases. One of the major clinical hurdles for the routine implementation of stem cell therapies is poor cell retention and survival, demanding the need to better understand these mechanisms while providing precise and scalable approaches to monitor these cell-based therapies in both pre-clinical and clinical scenarios. This poses significant multidisciplinary challenges regarding planning, defining the methodology and requirements, prototyping and different stages of testing. Aiming toward an optogenetic neural stem cell implant controlled by a smart wireless electronic frontend, we show how an iterative development methodology coupled with a modular design philosophy can mitigate some of these challenges. In this study, we present a miniaturized, wireless-controlled, modular multisensor platform with fully interfaced electronics featuring three different modules: an impedance analyzer, a potentiostat and an optical stimulator. We show the application of the platform for electrical impedance spectroscopy-based cell monitoring, optical stimulation to induce dopamine release from optogenetically modified neurons and a potentiostat for cyclic voltammetry and amperometric detection of dopamine release. The multisensor platform is designed to be used as an opto-electric headstage for future in vivo animal experiments.
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| 7. |
- Czolkos, Ilja, et al.
(author)
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Prediction of wastewater quality using amperometric bioelectronic tongues
- 2016
-
In: Biosensors & bioelectronics. - : Elsevier. - 0956-5663 .- 1873-4235. ; 75, s. 375-382
-
Journal article (peer-reviewed)abstract
- Wastewater samples from a Swedish chemi-thermo-mechanical pulp (CTMP) mill collected at different purification stages in a wastewater treatment plant (WWTP) were analyzed with an amperometric enzyme-based biosensor array in a flow-injection system. In order to resolve the complex composition of the wastewater, the array consists of several sensing elements which yield a multidimensional response. We used principal component analysis (PCA) to decompose the array's responses, and found that wastewater with different degrees of pollution can be differentiated. With the help of partial least squares regression (PLS-R), we could link the sensor responses to the Microtox (R) toxicity parameter, as well as to global organic pollution parameters (COD, BOD, and TOC). From investigating the influences of individual sensors in the array, it was found that the best models were in most cases obtained when all sensors in the array were included in the PLS-R model. We find that fast simultaneous determination of several global environmental parameters characterizing wastewaters is possible with this kind of biosensor array, in particular because of the link between the sensor responses and the biological effect onto the ecosystem into which the wastewater would be released. In conjunction with multivariate data analysis tools, there is strong potential to reduce the total time until a result is yielded from days to a few minutes. (C) 2015 Elsevier B.V. All rights reserved.
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| 8. |
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| 9. |
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| 10. |
- Davidsson, Richard, et al.
(author)
-
Microfluidic biosensing systems - Part I. Development and optimisation of enzymatic chemiluminescent mu-biosensors based on silicon microchips
- 2004
-
In: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0189. ; 4:5, s. 481-487
-
Journal article (peer-reviewed)abstract
- Chemiluminescent (CL) enzyme-based flow-through microchip biosensors (mu-biosensors) for detection of glucose and ethanol were developed for the purpose of monitoring real-time production and release of glucose and ethanol from microchip immobilised yeast cells. Part I of this study focuses on the development and optimisation of the mu-biosensors in a microfluidic sequential injection analysis (muSIA) system. Glucose oxidase (GOX) or alcohol oxidase (AOX) was co-immobilised with horseradish peroxidase (HRP) on porous silicon flow through microchips. The hydrogen peroxide ;produced from oxidation of the corresponding analyte (glucose or ethanol) took part in the chemiluminescent (CL) oxidation of luminol catalysed by HRP enhanced by addition of p-iodophenol ( PIP). All steps in the mSIA system, including control of syringe pump, multiposition valve (MPV) and data readout, were computer controlled. The influence of flow rate and luminol- and PIP concentration were investigated using a 2(3)-factor experiment using the GOX-HRP sensor. It was found that all estimated single factors and the highest order of interaction were significant. The optimum was found at 250 muM luminol and 150 muM PIP at a flow rate of 18 mul min(-1), the latter as a compromise between signal intensity and analysis time. Using the optimised system settings one sample was processed within 5 min. Two different immobilisation chemistries were investigated for both m-biosensors based on 3-aminopropyltriethoxsilane (APTS)- or polyethylenimine (PEI) functionalisation followed by glutaraldehyde (GA) activation. GOX-HRP mu-biosensors responded linear in a log-log format within the range 10-1000 mM glucose. Both had an operational stability of at least 8 days, but the PEI-GOX-HRP sensor was more sensitive. The AOX-HRP mu-biosensors responded linear (log-log) in the range between 1 and 10 mM ethanol, but the PEI-AOX-HRP sensor was in general more sensitive. Both sensors had an operational stability of at least 8 h, but with a half-life of 2-3 days.
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| 11. |
- Davidsson, Richard, et al.
(author)
-
Microfluidic biosensing systems - Part II. Monitoring the dynamic production of glucose and ethanol from microchip-immobilised yeast cells using enzymatic chemiluminescent mu-biosensors
- 2004
-
In: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0189. ; 4:5, s. 488-494
-
Journal article (peer-reviewed)abstract
- A microfluidic flow injection (muFIA) system was employed for handling and monitoring of cell-released products from living cells immobilised on silicon microchips. The dynamic release of glucose and ethanol produced from sucrose by immobilised Saccharomyces cerevisiae cells was determined using microchip biosensors (mu-biosensors) with either co-immobilised glucose oxidase-horseradish peroxidase (GOX-HRP), or alcohol oxidase-horseradish peroxidase (AOX-HRP), catalysing a series of reactions ending up with chemiluminescence (CL) generated from HRP-catalysed oxidation of luminol in presence of p-iodophenol (PIP). The yeast cells were attached by first treating them with polyethylenimine (PEI) followed by adsorption to the microchip surface. The cell loss during assaying was evaluated qualitatively using scanning electron microscopy (SEM), showing that no cells were lost after 35 min liquid handling of the cell chip at 10 mul min(-1). The enzymes were immobilised on microchips via PEI-treatment followed by glutaraldehyde (GA) activation. The GOX-HRP mu-biosensors could be used during five days without any noticeable decrease in response, while the AOX-HRP mu-biosensors showed continuously decreasing activity, but could still be used employing calibration correction. The glucose and ethanol released from the immobilised yeast chips were quantitatively monitored, by varying the incubation time with sucrose, showing the possibilities and advantages of using a microfluidic system set-up for cell-based assays.
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| 12. |
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| 13. |
- Dock, Eva, et al.
(author)
-
A steady-state and flow-through cell for screen-printed eight-electrode arrays
- 2005
-
In: Analytica Chimica Acta. - : Elsevier BV. - 1873-4324 .- 0003-2670. ; 531:2, s. 165-172
-
Journal article (peer-reviewed)abstract
- An electrochemical cell has been developed enabling amperometric steady-state- and flow-injection measurements with screen-printed arrays consisting of eight working electrodes (circle divide = 1 mm) arranged radially around a printed Ag/AgCl reference electrode in the centre. The cell contained a rotator, providing similar hydrodynamics over all the working electrodes in the array, which was manually centered under the rotator. The reproducibility of steady-state measurements with eight-electrode platinum or gold arrays in this cell was studied by measuring and comparing currents from ferricyanide reduction at each electrode in the array. It was found that the relative standard deviation (R.S.D.) for the currents at different electrodes on one array was below 5%. Similar R.S.D. was found if measurements were compared between several arrays. This indicates that manual insertion/positioning of the eight-electrode array in the cell and hydrodynamics at the electrodes provided measurement reproducibility similar to the reproducibility of manufacturing eight-electrode platinum or gold arrays by screen-printing. A comparative study was performed between screen-printed and through mask sprayed carbon arrays. It was found that the reproducibility of the sprayed arrays was similar to that of the platinum or gold screen-printed arrays, with R.S.D. values below 6% regarding the variation between electrodes within the same array and the variation between different arrays. To enable flow-injection measurements, a tube (0.4 mm inner diameter) was inserted into a hole drilled through the centre of the steady-state cell rotator. This construction made it possible to inject the solution into the cell through the tube (not rotating), while the rotator was spinning over the eight-electrode array. It was found that this combination of flow-injection and mixing by a rotator provided a uniform current response over the array electrodes and that, at optimum conditions, the R.S.D. values between the eight electrodes in the array were nearly the same as in case of the steady-state measurements, i.e., below 5%. (C) 2004 Elsevier B.V. All rights reserved.
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| 14. |
- Dock, Eva, et al.
(author)
-
Multivariate data analysis of dynamic amperometric biosensor responses from binary analyte mixtures - application of sensitivity correction algorithms
- 2005
-
In: Talanta. - : Elsevier BV. - 1873-3573 .- 0039-9140. ; 65:2, s. 298-305
-
Journal article (peer-reviewed)abstract
- In this paper, it is demonstrated that a single-receptor biosensor can be used to quantitatively determine each analyte in binary Mixtures LIS in multivariate data analysis tools based on the dynamic responses received from flow injection peaks. Mixtures with different concentrations of two phenolic compounds, catechol and 4-chlorophenol, were measured with a graphite electrode modified with tyrosinase enzyme at an applied potential of -50 mV versus Ag/AgCl. A correction algorithm based on measurements of references in-between samples was applied to compensate for biosensor ageing as well as differences caused by deviations between biosensor preparations. After correction, the relative prediction errors with partial least squares regression (PLS-R) for catechol and 4-chlorophenol were 7.4 and 5.5%, respectively, using an analysis sequence measured on one biosensor. Additional validation mixtures of the two phenols were measured with a new biosensor, prepared with the same procedure but with a different batch of tyrosinase enzyme. Using the mixture responses for the first sensor as a calibration set in PLS-R. the relative prediction errors of the validation mixtures, after applying correction procedures. were 7.0% for catechol and 16.0% for 4-chlorophenol. These preliminary results indicate that by applying correction algorithms it could be possible to use less stable biosensors in continuous on-line measurements together with multivariate data analysis without time-consuming calibration procedures. (C) 2004 Elsevier B.V. All rights reserved.
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| 15. |
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| 16. |
- Esmail Tehrani, Sheida, et al.
(author)
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Hydrogen Peroxide Detection Using Prussian Blue-modified 3D Pyrolytic Carbon Microelectrodes
- 2021
-
In: Electroanalysis. - : John Wiley & Sons. - 1040-0397 .- 1521-4109. ; 33:12, s. 2516-2528
-
Journal article (peer-reviewed)abstract
- A highly sensitive amperometric Prussian blue-based hydrogen peroxide sensor was developed using 3D pyrolytic carbon microelectrodes. A 3D printed multielectrode electrochemical cell enabled simultaneous highly reproducible Prussian blue modification on multiple carbon electrodes. The effect of oxygen plasma pre-treatment and deposition time on Prussian blue electrodeposition was studied. The amperometric response of 2D and 3D sensors to the addition of hydrogen peroxide in mu M and sub-mu M concentrations in phosphate buffer was investigated. A high sensitivity comparable to flow injection systems and a detection limit of 0.16 mu M was demonstrated with 3D pyrolytic carbon microelectrodes at stirred batch condition
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| 17. |
- Fiorenzano, Alessandro, et al.
(author)
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Single-cell transcriptomics captures features of human midbrain development and dopamine neuron diversity in brain organoids
- 2021
-
In: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 12:1
-
Journal article (peer-reviewed)abstract
- Three-dimensional brain organoids have emerged as a valuable model system for studies of human brain development and pathology. Here we establish a midbrain organoid culture system to study the developmental trajectory from pluripotent stem cells to mature dopamine neurons. Using single cell RNA sequencing, we identify the presence of three molecularly distinct subtypes of human dopamine neurons with high similarity to those in developing and adult human midbrain. However, despite significant advancements in the field, the use of brain organoids can be limited by issues of reproducibility and incomplete maturation which was also observed in this study. We therefore designed bioengineered ventral midbrain organoids supported by recombinant spider-silk microfibers functionalized with full-length human laminin. We show that silk organoids reproduce key molecular aspects of dopamine neurogenesis and reduce inter-organoid variability in terms of cell type composition and dopamine neuron formation.
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| 18. |
- Heiskanen, Arto, et al.
(author)
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Amperometric monitoring of redox activity in living yeast cells: comparison of menadione and menadione sodium bisulfite as electron transfer mediators
- 2004
-
In: Electrochemistry Communications. - : Elsevier BV. - 1388-2481. ; 6:2, s. 219-224
-
Journal article (peer-reviewed)abstract
- An amperometric method was applied for real-time monitoring of intracellular redox enzyme activity. Baker’s yeast (Saccharomyces cerevisiae) cells were immobilized on platinum microband electrodes and mediated anodic currents were measured. The currents were observed in the absence and in the presence of glucose as a source of reducing equivalents, NADH and NADPH. 2-Methyl-1,4-naphthoquinone (menadione, vitamin K3) and water soluble 2-methyl-1,4-naphthoquinone sodium bisulfite (menadione sodium bisulfite MSB) were compared as artificial electron acceptors for their ability to transduce internal cellular redox activity into electrode current. It was found that hydrophobic menadione was superior to its water-soluble bisulfite derivative for probing whole intact cells.
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| 19. |
- Heiskanen, Arto, et al.
(author)
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Bioelectrochemical probing of intracellular redox processes in living yeast cells-application of redox polymer wiring in a microfluidic environment
- 2013
-
In: Analytical and Bioanalytical Chemistry. - : Springer Science and Business Media LLC. - 1618-2642 .- 1618-2650. ; 405:11, s. 3847-3858
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Journal article (peer-reviewed)abstract
- Conventionally, microbial bioelectrochemical assays have been conducted using immobilized cells on an electrode that is placed in an electrochemical batch cell. In this paper, we describe a developed microfluidic platform with integrated microelectrode arrays for automated bioelectrochemical assays utilizing a new double mediator system to map redox metabolism and screen for genetic modifications in Saccharomyces cerevisiae cells. The function of this new double mediator system based on menadione and osmium redox polymer (PVI-Os) is demonstrated. "Wiring" of S. cerevisiae cells using PVI-Os shows a significant improvement of bioelectrochemical monitoring in a microfluidic environment and functions as an effective immobilization matrix for cells that are not strongly adherent. The function of the developed microfluidic platform is demonstrated using two strains of S. cerevisiae, ENY. WA and its deletion mutant EBY44, which lacks the enzyme phosphoglucose isomerase. The cellular responses to introduced glucose and fructose were recorded for the two S. cerevisiae strains, and the obtained results are compared with previously published work when using an electrochemical batch cell, indicating that microfluidic bioelectrochemical assays employing the menadione-PVI-Os double mediator system provides an effective means to conduct automated microbial assays.
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| 20. |
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| 21. |
- Heiskanen, Arto, et al.
(author)
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Mediator-assisted simultaneous probing of cytosolic and mitochondrial redox activity in living cells.
- 2009
-
In: Analytical Biochemistry. - : Elsevier BV. - 1096-0309 .- 0003-2697. ; 384:1, s. 11-19
-
Journal article (peer-reviewed)abstract
- This work describes an electron transfer mediator-assisted amperometric flow injection method for assessing redox enzyme activity in different subcellular compartments of the phosphoglucose isomerase deletion mutant strain of Saccharomyces cerevisiae, EBY44. The method is demonstrated using the ferricyanide-menadione double mediator system to study the effect of dicoumarol, an inhibitor of cytosolic and mitochondrial oxidoreductases and an uncoupler of the electron transport chain. Evaluation of the role of NAD(P)H-producing pathways in mediating biological effects is facilitated by introducing either fructose or glucose as the carbon source, yielding either NADH or NADPH through the glycolytic or pentose phosphate pathway, respectively. Respiratory noncompetent cells show greater inhibition of cytosolic menadione-reducing enzymes when NADH rather than NADPH is produced. Spectrophotometric in vitro assays show no difference between the cofactors. Respiratory competent cells show cytosolic inhibition only when NADPH is produced, whereas production of NADH reveals uncoupling at low dicoumarol concentrations and inhibition of complexes III and IV at higher concentrations. Spectrophotometric assays only indicate the presence of cytosolic inhibition regardless of the reduced cofactor used. This article shows the applicability of the amperometric method and emphasizes the significance of determining biological effects of chemicals in living cells.
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| 22. |
- Heiskanen, Arto, et al.
(author)
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Monitoring of Saccharomyces cerevisiae Cell Proliferation on Thiol-Modified Planar Gold Microelectrodes Using Impedance Spectroscopy.
- 2008
-
In: Langmuir. - : American Chemical Society (ACS). - 0743-7463 .- 1520-5827. ; 24:16, s. 9066-9073
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Journal article (peer-reviewed)abstract
- An impedance spectroscopic study of the interaction between thiol-modified Au electrodes and Saccharomyces cerevisiae of strain EBY44 revealed that the cells formed an integral part of the interface, modulating the capacitive properties until a complete monolayer was obtained, whereas the charge transfer resistance ( R ct) to the redox process of [Fe(CN)6] (3-/4-) showed a linear relationship to the number of cells even beyond the monolayer coverage. R ct showed strong pH dependence upon increasing the pH of the utilized buffer to 7.2. Upon addition of S. cerevisiae cells at pH 7.2, the obtained value of R ct showed over 560% increase with respect to the value obtained on the same thiol-modified electrode without cells. It was demonstrated that real-time monitoring of S. cerevisiae proliferation, with frequency-normalized imaginary admittance (real capacitance) as the indicator, was possible using a miniaturized culture system, ECIS Cultureware, with integrated planar cysteamine-modified Au microelectrodes. A monolayer coverage was reached after 20-28 h of cultivation, observed as an approximately 15% decrease in the real capacitance of the system.
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| 23. |
- Jain, Seema Rani, et al.
(author)
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A chemiluminescence flow immunosensor based on a porous monolithic metacrylate and polyethylene composite disc modified with Protein G
- 2004
-
In: Biosensors & Bioelectronics. - : Elsevier BV. - 1873-4235 .- 0956-5663. ; 19:8, s. 795-803
-
Journal article (peer-reviewed)abstract
- A generic, fast, sensitive and new type of flow immunosensor has been developed. The basis is a monolithic porous poly(glycidyl methacrylate-co-trimethylolpropane trimethacrylate) polymer disc modified with protein G, placed in a fountain type flow cell compartment, in close proximity to a photomultiplier tube (PMT). Analyte and HRP labelled analyte derivative (tracer) compete for anti-analyte antibody binding sites. The mixture is then injected into the flow immunosensor system where the formed analyte- and tracer-antibody complexes are trapped by the monolithic protein G disc. The amount of bound tracer, inversely related to the concentration of analyte in the sample, is determined in a second step by injection of luminol, p-iodophenol and H2O2, generating enhanced chemiluminescence (CL) with horseradish peroxidase (HRP). A third and final step is need for regeneration of the protein G disc so that a new analysis cycle can take place. The performance of the disc immunosensor system was compared with a one step continuous flow injection immunoassay (FIIA) system, using the same reagents and a protein G column, in terms of assay sensitivity and influence of matrix effects from various water samples (millipore-, tap- and surface water). The detection limit for the analyte atrazine in PBS and surface water (SW) was 0.208±0.004 g l−1 (PBS) and 0.59±0.120 g l−1 (SW) for the FIIA and 0.033±0.003 g l−1 (PBS) and 0.038±0.003 g l−1 (SW) for the disc immunosensor. Statistical comparison of the two systems shows that the disc immunosensor results were significantly less influenced by the sample matrix, which is explained by the fact that the sample in the FIIA arrives simultaneously with the matrix to the detector, whereas these are separated in time in the disc immunosensor system.
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| 24. |
- Kajtez, Janko, et al.
(author)
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3D biomaterial models of human brain disease
- 2021
-
In: Neurochemistry International. - : Elsevier BV. - 0197-0186. ; 147
-
Journal article (peer-reviewed)abstract
- Inherent limitations of the traditional approaches to study brain function and disease, such as rodent models and 2D cell culture platforms, have led to the development of 3D in vitro cell culture systems. These systems, products of multidisciplinary efforts encompassing stem cell biology, materials engineering, and biofabrication, have quickly shown great potential to mimic biochemical composition, structural properties, and cellular morphology and diversity found in the native brain tissue. Crucial to these developments have been the advancements in stem cell technology and cell reprogramming protocols that allow reproducible generation of human subtype-specific neurons and glia in laboratory conditions. At the same time, biomaterials have been designed to provide cells in 3D with a microenvironment that mimics functional and structural aspects of the native extracellular matrix with increasing fidelity. In this article, we review the use of biomaterials in 3D in vitro models of neurological disorders with focus on hydrogel technology and with biochemical composition and physical properties of the in vivo environment as reference.
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| 25. |
- Kajtez, Janko, et al.
(author)
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3D-Printed Soft Lithography for Complex Compartmentalized Microfluidic Neural Devices
- 2020
-
In: Advanced Science. - : Wiley. - 2198-3844. ; 7:16
-
Journal article (peer-reviewed)abstract
- Compartmentalized microfluidic platforms are an invaluable tool in neuroscience research. However, harnessing the full potential of this technology remains hindered by the lack of a simple fabrication approach for the creation of intricate device architectures with high-aspect ratio features. Here, a hybrid additive manufacturing approach is presented for the fabrication of open-well compartmentalized neural devices that provides larger freedom of device design, removes the need for manual postprocessing, and allows an increase in the biocompatibility of the system. Suitability of the method for multimaterial integration allows to tailor the device architecture for the long-term maintenance of healthy human stem-cell derived neurons and astrocytes, spanning at least 40 days. Leveraging fast-prototyping capabilities at both micro and macroscale, a proof-of-principle human in vitro model of the nigrostriatal pathway is created. By presenting a route for novel materials and unique architectures in microfluidic systems, the method provides new possibilities in biological research beyond neuroscience applications.
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| 26. |
- Kajtez, Janko, et al.
(author)
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Embedded 3D Printing in Self-Healing Annealable Composites for Precise Patterning of Functionally Mature Human Neural Constructs
- 2022
-
In: Advanced science (Weinheim, Baden-Wurttemberg, Germany). - : Wiley. - 2198-3844. ; 9:25
-
Journal article (peer-reviewed)abstract
- Human in vitro models of neural tissue with tunable microenvironment and defined spatial arrangement are needed to facilitate studies of brain development and disease. Towards this end, embedded printing inside granular gels holds great promise as it allows precise patterning of extremely soft tissue constructs. However, granular printing support formulations are restricted to only a handful of materials. Therefore, there has been a need for novel materials that take advantage of versatile biomimicry of bulk hydrogels while providing high-fidelity support for embedded printing akin to granular gels. To address this need, Authors present a modular platform for bioengineering of neuronal networks via direct embedded 3D printing of human stem cells inside Self-Healing Annealable Particle-Extracellular matrix (SHAPE) composites. SHAPE composites consist of soft microgels immersed in viscous extracellular-matrix solution to enable precise and programmable patterning of human stem cells and consequent generation mature subtype-specific neurons that extend projections into the volume of the annealed support. The developed approach further allows multi-ink deposition, live spatial and temporal monitoring of oxygen levels, as well as creation of vascular-like channels. Due to its modularity and versatility, SHAPE biomanufacturing toolbox has potential to be used in applications beyond functional modeling of mechanically sensitive neural constructs.
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| 27. |
- Khampha, Wanida, et al.
(author)
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Specific detection of L-glutamate in food using flow-injection analysis and enzymatic recycling of substrate
- 2004
-
In: Analytica Chimica Acta. - : Elsevier BV. - 1873-4324 .- 0003-2670. ; 518:1-2, s. 127-135
-
Journal article (peer-reviewed)abstract
- A flow injection analysis (FIA) system for specific determination of L-glutamate in food samples based on a bi-enzymatic amplification system has been developed. The content of L-glutamate in the sample was amplified by cycling between L-glutamate dehydrogenase (GIDH) and a novel enzyme, D-phenylglycine aminotransferase (D-PhgAT). In this system, GIDH converts L-glutamate to 2-oxoglutarate with concomitant reduction of NAD(+) to NADH. D-PhgAT transfers an amino group from D-4-hydroxyphenylglycine to 2-oxoglutarate, thus recycling L-glutamate. Accumulation of NADH in the course of the enzymatic recycling was monitored both by fluorescence and UV absorbance and used for quantification of L-glutamate. The assay was characterized by high long-term stability (at least 70 days) and good reproducibility (within-day and between-day RSDs were 4.3-7.3% and 8.9%). The fluorimetric assay was slightly more sensitive with a L-glutamate detection limit of 0.4 muM and linear range of 2.5-50 muM. The assay was specific for L-glutamate, with recoveries between 95-103% in the presence of 17 different amino acids tested one by one. The method was applied to analysis of real food samples and results were correlated with a commercial Boehringer Mannheim assay kit. (C) 2004 Elsevier B.V. All rights reserved.
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| 28. |
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| 29. |
- Kostesha, Natalie, et al.
(author)
-
Probing the redox metabolism in the strictly anaerobic, extremely thermophilic, hydrogen-producing Caldicellulosiruptor saccharolyticus using amperometry
- 2011
-
In: Extremophiles. - : Springer Science and Business Media LLC. - 1433-4909 .- 1431-0651. ; 15:1, s. 77-87
-
Journal article (peer-reviewed)abstract
- Changes in the redox metabolism in the anaerobic, extremely thermophilic, hydrogen-forming bacterium Caldicellulosiruptor saccharolyticus were probed for the first time in vivo using mediated amperometry with ferricyanide as a thermotolerant external mediator. Clear differences in the intracellular electron flow were observed when cells were supplied with different carbon sources. A higher electrochemical response was detected when cells were supplied with xylose than with sucrose or glucose. Moreover, using the mediated electrochemical method, it was possible to detect differences in the electron flow between cells harvested in the exponential and stationary growth phases. The electron flow of C. saccharolyticus was dependent on the NADH- and reduced ferredoxin generation flux and the competitive behavior of cytosolic and membrane-associated oxidoreductases. Sodium oxamate was used to inhibit the NADH-dependent lactate dehydrogenase, upon which more NADH was directed to membrane-associated enzymes for ferricyanide reduction, leading to a higher electrochemical signal. The method is noninvasive and the results presented here demonstrate that this method can be used to accurately detect changes in the intracellular electron flow and to probe redox enzyme properties of a strictly anaerobic thermophile in vivo.
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| 30. |
- Kostesha, Natalie, et al.
(author)
-
Real-time detection of cofactor availability in genetically modified living Saccharomyces cerevisiae cells - Simultaneous probing of different geno- and phenotypes.
- 2009
-
In: Bioelectrochemistry. - : Elsevier BV. - 1878-562X .- 1567-5394. ; 76, s. 180-188
-
Journal article (peer-reviewed)abstract
- This work describes a mediated amperometric method for simultaneous real-time probing of the NAD(P)H availability in two different phenotypes, fermentative and respiratory, of the phosphoglucose isomerase deletion mutant strain of S. cerevisiae, EBY44 [ENY.WA-1A pgi1-1D::URA3], and its parental strain, ENY.WA-1A. The developed method is based on multichannel detection using microelectrode arrays. Its versatility was demonstrated by using four microelectrode arrays for simultaneously monitoring the NAD(P)H availability of both geno- and phenotypes under the influence of two different carbon sources, glucose and fructose, as well as the cytosolic and mitochondrial inhibitor and uncoupler, dicoumarol. The obtained results indicate that the method is capable of accurately and reproducibly (overall relative standard error of mean 3.2%) mapping the real-time responses of the cells with different genotype-phenotype combinations. The ENY.WA cells showed the same response to glucose and fructose when dicoumarol was used; fermentative cells indicated the presence of cytosolic inhibition and respiratory cells a net effect of mitochondrial uncoupling. EBY44 cells showed cytosolic inhibition with the exception of respiratory cells when fructose was used as carbon source.
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| 31. |
- Krikstolaityte, Vida, et al.
(author)
-
Development of a plastic membrane containing micro-hole(s) for a potential bio-sensing application
- 2017
-
In: Procedia Technology. - : Elsevier. - 2212-0173. ; 27:Special Issue Biosensors 2016, s. 252-253
-
Journal article (peer-reviewed)abstract
- In this work, a poly (methyl methacrylate) membrane containing micro-holes (MHs) as a prototype of a simple sensing platform of a lab-on-a-chip device has been developed for a potential analysis of clinical fluidic samples. A four probe electrochemical impedance spectroscopy (EIS) setup, with two electrodes placed on each side of the membrane, was adopted for monitoring the MH impedance (Fig. 1a). The setup was used to investigate, if EIS is suitable to sense the trapping of an analyte inside the MHs. Latex micro-beads with a diameter of 10 mu m were used to test clogging of the MHs. Additionally, finite element model simulations were performed using Comsol Multiphysics software to theoretically evaluate the sensitivity field of the EIS measurement along the MHs. (C) 2017 The Authors. Published by Elsevier Ltd.
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| 32. |
- Lutz, Mareike, 1967-, et al.
(author)
-
Effects of different additives on a tyrosinase based carbon paste electrode
- 1995
-
In: Analytica Chimica Acta. - Amsterdam : Elsevier. - 0003-2670 .- 1873-4324. ; 305:1-3, s. 8-17
-
Journal article (peer-reviewed)abstract
- The influence of a number of solid and chemical additives on the sensitivity and operational stability of a tyrosinase carbon paste electrode was studied. Cyclic voltammograms were run of the electrochemically active catechol/o-quinone couple on unmodified and additive modified carbon paste electrodes without tyrosinase. This was done in order to study the influence of these additives on the pure electrochemistry of the carbon paste. The influence on the total system (additive and enzyme modified carbon paste electrode) was studied in the flow injection mode. In some instances a dramatic improvement of the direct electron transfer of the catechol/o-quinone couple was obtained with both solid and chemical additives included in the carbon paste. A similar improvement of biosensor sensitivity in the flow injection mode was obtained with most chemical additives whereas the solid additives had a negative impact on biosensor sensitivity. The results obtained in this work indicate that these additives influence the purely electrochemical processes at the carbon paste and/or the performance of the enzyme in the carbon paste environment. How and why these additives can possibly influence the biosensor performance are discussed. © 1995.
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| 33. |
- Marko-Varga, György, et al.
(author)
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Effect of HY-Zeolites on the Performance of Tyrosinase-Modified Carbon Paste Electrodes
- 1996
-
In: Electroanalysis. - Weinheim : Wiley-VCH Verlagsgesellschaft. - 1040-0397 .- 1521-4109. ; 8:12, s. 1121-1126
-
Journal article (peer-reviewed)abstract
- The dependence of electrode response on additive properties in enzyme-modified carbon paste was studied. Four different HY-zeolite powders, dealuminated to different extents and characterized by both Si/Al ratio and hydrophilicity, were used as the carbon paste modifiers. The enzyme tyrosinase used in biosensors for the detection of catechol and other phenolic compounds was chosen as the model system for the construction of a composite carbon paste biosensor incorporating different HY-zeolites as additives. Tyrosinase was trapped on the HY-zeolite particles from a buffer solution, dried and mixed with graphite powder and a pasting oil. It was found that by incorporating HY-zeolites into the carbon paste the heterogeneous reaction rate of catechol redox conversion and the signal response for catechol were increased. In the latter case a higher response was observed for increased hydrophilicity, i.e., decreased Si/Al ratio of the HY-zeolite. The carbon paste/solution interface is considered to be an aqueous/organic phase and the characteristics of the enzyme-modified carbon paste electrode are related to theories, explaining enzymatic catalysis in organic solvents.
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| 34. |
- Mie, Axel, et al.
(author)
-
Analysis of triazines and associated metabolites with electrospray ionization field-asymmetric ion mobility spectrometry/mass spectrometry.
- 2008
-
In: Analytical Sciences. - 0910-6340. ; 24:8, s. 973-978
-
Journal article (peer-reviewed)abstract
- Triazines comprise an important pollutant class owing to continued use in certain countries, and owing to strong environmental persistence that leads to problems even in countries like Sweden where the use of triazines has been prohibited for some years. We investigated mass-selective detection for analysis of triazines. More specifically, we studied the background reduction and sensitivity enhancement that result from the use of a new interface technique, field-asymmetric ion mobility spectrometry (FAIMS), in conjunction with electrospray ionization ion-trap mass spectrometry. This technique allows for ion sorting and discrimination against the considerable "chemical noise", nonspecific cluster and fragment ions, which are typically generated in electrospray ionization. This paper presents results of a pilot study of triazines and some metabolites in ideal solvents. Our long-range goal is automated analysis with mass-selective detection coupled to membrane-based sample cleanup and enrichment for additional enhancement in sensitivity.
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| 35. |
- Nistor, Catalin, et al.
(author)
-
A capillary-based amperometric flow immunoassay for 2,4,6-trichlorophenol
- 2003
-
In: Analytical and Bioanalytical Chemistry. - : Springer Science and Business Media LLC. - 1618-2642. ; 375:1, s. 125-132
-
Journal article (peer-reviewed)abstract
- This paper describes the development of two different capillary-based heterogeneous competitive flow immunoassay formats (capillary flow injection immunoassay (CFIIA) and capillary sequential injection immunoassay (CSIIA)) for the determination of 2,4,6-trichlorophenol (2,4,6-TCP). The assays are based on the competition between the analyte and an analyte derivative labelled with the enzyme #-galactosidase, for an anti-TCP antibody, followed by the injection of the mixture at equilibrium into a flow stream, where separation between the fractions bound and unbound to the antibody is performed in a glass capillary containing immobilised protein A. The antibody-tracer fraction retained inside the protein A capillary was measured by injection of 4-aminophenyl-#-D-galactoside (4-APG), followed by amperometric detection of the enzymatically generated 4-aminophenol (4-AP), leading to a negative correlation between the signal and the analyte concentration.
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| 36. |
- Nistor, Catalin, et al.
(author)
-
A glucose dehydrogenase biosensor as an additional signal amplification step in an enzyme-flow immunoassay.
- 2002
-
In: Analyst. - : Royal Society of Chemistry (RSC). - 1364-5528. ; 127:8, s. 1076-1081
-
Journal article (peer-reviewed)abstract
- Both the antibody affinity and the detectability of the label are essential in deciding the final characteristics of a heterogeneous immunoassay. This paper describes an approach to obtain a supplementary enhancement of the signal generated by using an enzyme label, e.g., by including the product of the enzymatic reaction in an additional amplification cycle during the detection step performed with an amperometric biosensor based on glucose dehydrogenase (GDH). An immunoassay format with a labelled analyte derivative that competes with the analyte present in the sample for a limited amount of antibody binding sites was employed. The beta-galactosidase label hydrolyses the substrate aminophenyl-beta-galactopyranoside, and the generated aminophenol enters then into a bioelectrocatalytic amplification cycle at the GDH biosensor. The principle was applied for determination of 4-nitrophenol, with the best minimal concentration of 1.5 microM and a midpoint of the calibration of 24 microM. The potentials and limitations of such a system are discussed.
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| 37. |
- Nistor, Catalin, et al.
(author)
-
Detection of Escherichia coli in water by culture-based amperometric and luminometric methods.
- 2002
-
In: Water Science and Technology. - 0273-1223. ; 45:4-5, s. 191-199
-
Journal article (peer-reviewed)abstract
- The application of amperometric biosensor- and chemiluminiscence based methods for rapid detection of viable E. coli in water has been investigated. An amplification of the amperometric signal by a factor of 4 was obtained when the cellobiose dehydrogenase (CDH) biosensor was used instead of a plain graphite electrode for detection of b-galactosidase (b-GAL) activity at 22.5 degrees C. A linear correlation was demonstrated for detection time (DT) vs. initial concentrations (logarithmic units) of E. coli IT1 and E. coli in environmental samples, respectively, by use of the CDH biosensor or a chemiluminometric technique. The study has shown that an E. coli concentration > or = 10(4) cfu/100 mL in environmental samples was determined by the CDH biosensor within one working day. However, further reduction of the DT can be obtained, e.g. by increasing the signal amplification factor using other biosensors.
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| 38. |
- Nistor, Catalin, et al.
(author)
-
In-field monitoring of cleaning efficiency in waste water treatment plants using two phenol-sensitive biosensors
- 2002
-
In: Analytica Chimica Acta. - 1873-4324. ; 456:1, s. 3-17
-
Journal article (peer-reviewed)abstract
- Two amperometric biosensors based on the enzymes cellobiose dehydrogenase (CDH) and quinoprotein-dependent glucose dehydrogenase (GDH), have been applied for monitoring the phenolic content in water samples, collected at different stages of a waste water treatment process, thus representing different cleaning levels of two waste water treatment plants (WWTPs). The biosensor measurements were performed in-field, compared with the results obtained by liquid chromatography-mass spectrometry and were further correlated with the cleaning efficiencies of the WWTPs. The effect of several potentially interfering compounds on the sensor response was also studied. The general purpose of the study was to evaluate the potential use of biosensors, not as quantitative tools for phenol analysis, but rather as screening tools indicating a certain trend, i.e. compounds present or not present, and potential correlation with sample toxicity. It was found that the biosensors and LC-MS results were not quantitatively comparable, however, both sensors could follow the decrease of the phenol content from the influent, primary treated and effluent waters. In addition, the correlation between biosensor inhibition and sample toxicity is discussed.
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| 39. |
- Nistor, Catalin, et al.
(author)
-
Multivariate analysis to separate the signal given by cross-reactants in immunoassay with sample matrix dilution
- 2004
-
In: Analytical and Bioanalytical Chemistry. - : Springer Science and Business Media LLC. - 1618-2642 .- 1618-2650. ; 380:7-8, s. 898-907
-
Journal article (peer-reviewed)abstract
- This paper describes a new approach to achieve selectivity in an immunoassay by separating the signals given by two cross-reactive compounds present simultaneously in a complex sample matrix. The method is based on the sequential dilution of the sample containing a mixture of the two analytes, spiking each diluted sample with a reference compound, and the detection by enzyme-linked immunosorbent assay (ELISA). The obtained multivariate response was used for the individual calibrations of the assay for each. of the two cross-reactants simultaneously by using principal component analysis (PCA) and partial least squares regression (PLSR) data modeling. The calibration models showed. that the signal separation due the analytes 2,4-dinitro-phenol (2,4-DNP) and 4-nitrophenol (4-NP) was possible with a prediction concentration error of 1.4 muM and 72 muM, respectively.
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| 40. |
- Pupinyo, Naricha, et al.
(author)
-
Impedimetric melanoma invasion assay device using a simple paper membrane and stencil-printed electrode on PMMA substrate
- 2020
-
In: Sensing and Bio-Sensing Research. - : Elsevier BV. - 2214-1804. ; 29
-
Journal article (peer-reviewed)abstract
- The transwell assay is currently the most popular approach to studying cellular invasion due to its ease of use and readout, and the possibility for quantitative measurements. However, it only allows end-point measurements without the possibility for real-time tracking of the dynamics of cell movement during an invasion. Moreover, it requires cell labeling, and construction of customized devices is hampered by the commercial standard membrane inserts, only available in certain designs. Recently, paper has been used as a scaffold for three-dimensional (3D) cell cultures. Because of its microfibrous structure and easy handling, it could be a versatile alternative as a membrane insert in customized devices. Here, we develop a low-cost real-time invasion assay device using paper as an alternative membrane insert. The device was designed for two-electrode impedance measurements and fabricated using CNC micromilling. It also comprised a disposable low-cost stencil-printed working electrode on a poly(methyl methacrylate) substrate below the membrane and glassy carbon counter electrode above the membrane inserted in a specially designed lid. Thus, the impedance measurements during cell invasion addressed the entire membrane. We demonstrated the function of the device by monitoring the invasion of B16 melanoma 4A5 cells from a mouse using insulin growth factor-1 as the chemoattractant. The cell invasion on paper was visualized using scanning electron microscopy and confocal microscopy with Z-stack 3D imaging. Melanoma cell invasion could be observed within 7 h after the chemoattractant treatment, which was faster than the conventional assay and less likely to be influenced by cell proliferation.
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| 41. |
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| 42. |
- Rose, A, et al.
(author)
-
GDH biosensor based off-line capillary immunoassay for alkylphenols and their ethoxylates
- 2002
-
In: Biosensors & Bioelectronics. - 1873-4235. ; 17:11-12, s. 1033-1043
-
Journal article (peer-reviewed)abstract
- The application of a quinoprotein glucose dehydrogenase modified thick-film sensor as label detector in a capillary immunoassay (CIA) for xenoestrogens is presented. The detection of the alkylphenols and their ethoxylates is based on the competition between the analyte and tracer molecules for the binding sites of anti-alkylphenol ethoxylate antibodies. This assay is performed off-line in small disposable PVC capillaries coated with immobilized antibodies. This format allows the combination of the assay with a small portable device potentially useful for on-site environmental monitoring. Beside high amplification the utilization of beta-galactosidase as enzyme label allows the direct combination with a GDH biosensor at optimal pH conditions. The bioelectrocatalytic properties of this biosensor offer an additional amplification and thus allow a very sensitive quantification of 4-aminophenol, generated by the beta-galactosidase. Detection limits of the analytes in the mug/l range were obtained, while other phenolics and surfactants showed no or very little cross reactivity.
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| 43. |
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| 44. |
- Sapelnikova, Svetlana, et al.
(author)
-
Amperometric sensors based on tyro sinase-modified screenprinted arrays
- 2003
-
In: Talanta. - 1873-3573. ; 61:4, s. 473-483
-
Journal article (peer-reviewed)abstract
- This paper describes the design, development and characteristics of a tyrosinase (polyphenol oxidase) modified amperometric screen-printed biosensor array, with the enzyme cross-linked in a redox-hydrogel namely the PVI13-dmeOs polymer. Two types of Au-screen-printed four-channel electrode arrays, differing in design and insulating layer, were compared and investigated. Au-, graphite-coated-Au- and Carbopack C-coated-Au-surfaces, serving as the basis for tyrosinase immobilisation, were investigated and the performances of the different arrays were evaluated and compared in terms of their electrocatalytic characteristics, as well as operational- and storage stability using catechol as model substrate. It was found that the Carbopack C-coated array was the best choice for tyrosinase immobilisation procedure mainly due to a higher mechanical stability of the deposited enzyme layer, combined with good sensitivity and stability for up to 6 months of use. In the batch mode the biosensors responded linearly to catechol up to 30 muM with limits of detection from 0.14 muM. Parameters from cyclic voltammograms indicated that the reversibility of the direct electrochemical reaction for catechol on the three types of electrode surfaces (no tyrosinase modification) was not the limiting factor for the construction and performance of tyrosinase biosensors. (C) 2003 Elsevier B.V. All rights reserved.
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| 45. |
- Sapelnikova, Svetlana, et al.
(author)
-
Screen-printed multienzyme arrays for use in amperometric batch and flow systems
- 2003
-
In: Analytical and Bioanalytical Chemistry. - : Springer Science and Business Media LLC. - 1618-2642 .- 1618-2650. ; 376:7, s. 1098-1103
-
Journal article (peer-reviewed)abstract
- Screen-printing technology for electrode fabrication enables construction of amperometric devices suitable for combination of several enzyme electrodes. To develop a biosensor array for characterisation of wastewaters, tyrosinase and horseradish peroxidase (HRP) or cholinesterase-modified electrodes were combined on the same array. The behaviour of the tyrosinase-modified electrode in the presence of hydrogen peroxide (required co-substrate for the HRP-modified electrode) and acetylthiocholine chloride (required co-substrate for cholinesterase) was studied. Performance of bi-enzyme biosensor arrays in the batch mode and in the flow-injection system are discussed.
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| 46. |
- Simsa, Robin, et al.
(author)
-
Brain organoid formation on decellularized porcine brain ECM hydrogels
- 2021
-
In: PLOS ONE. - : Public Library of Science. - 1932-6203. ; 16:1
-
Journal article (peer-reviewed)abstract
- Human brain tissue models such as cerebral organoids are essential tools for developmental and biomedical research. Current methods to generate cerebral organoids often utilize Matrigel as an external scaffold to provide structure and biologically relevant signals. Matrigel however is a nonspecific hydrogel of mouse tumor origin and does not represent the complexity of the brain protein environment. In this study, we investigated the application of a decellularized adult porcine brain extracellular matrix (B-ECM) which could be processed into a hydrogel (B-ECM hydrogel) to be used as a scaffold for human embryonic stem cell (hESC)-derived brain organoids. We decellularized pig brains with a novel detergent- and enzyme-based method and analyzed the biomaterial properties, including protein composition and content, DNA content, mechanical characteristics, surface structure, and antigen presence. Then, we compared the growth of human brain organoid models with the B-ECM hydrogel or Matrigel controls in vitro. We found that the native brain source material was successfully decellularized with little remaining DNA content, while Mass Spectrometry (MS) showed the loss of several brain-specific proteins, while mainly different collagen types remained in the B-ECM. Rheological results revealed stable hydrogel formation, starting from B-ECM hydrogel concentrations of 5 mg/mL. hESCs cultured in B-ECM hydrogels showed gene expression and differentiation outcomes similar to those grown in Matrigel. These results indicate that B-ECM hydrogels can be used as an alternative scaffold for human cerebral organoid formation, and may be further optimized for improved organoid growth by further improving protein retention other than collagen after decellularization.
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| 47. |
- Skjolding, Lars Henrik, et al.
(author)
-
Characterisation of nano-interdigitated electrodes
- 2008
-
In: Journal of Physics: Conference Series. - : IOP Publishing. - 1742-6588 .- 1742-6596. ; 100, s. 052045-052045
-
Conference paper (peer-reviewed)abstract
- Interdigitated electrodes made up of two individually addressable interdigitated comb-like electrode structures have frequently been suggested as ultra sensitive electrochemical biosensors. Since the signal enhancement effects due to cycling of the reduced and oxidized species are strongly dependent on the inter electrode distances, since the nature of the enhancement is due to overlying diffusion layers, inter digitated electrodes with an electrode separation of less the non emicrometer a redesired for maximum signal amplification. Fabrication of submicron structures can only be made by advanced lithography techniques. By use of electron be amlithography we have fabricated arrays of interdigitated electrodes with an electrode separation distance of 200nm and an electrode finger width of likewise 200nm. The entire electrode structure is 100 micrometre times 100 micrometre, and the active electrode area is dictated by the opening in the passivation layer, that is defined by UV lithography. Here we report measurements of redox cycling of ferrocyanide by coupled cyclic voltammograms, where the potential atone of the working electrodes are varied and either an oxidising or reducing potential is applied to the complimentary interdigitated electrode. The measurements show fast conversion and high collection efficiency round 87% as expected for nano-interdigitated electrodes.
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| 48. |
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| 49. |
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| 50. |
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