Sungbo Cho

A capacitive biosensor based on an interdigitated electrode with nanoislands

A capacitive biosensor based on an interdigitated electrode (IDE) with nanoislands was developed for label-free detection of antigen-antibody interactions. To enable sensitive capacitive detection of protein adsorption, the nanoislands were fabricated between finger electrodes of the IDE. The effect of the nanoislands on the sensitive capacitive measurement was estimated using horseradish peroxidase (HRP) as a model protein. Additionally, a parylene-A film was coated on the IDE with nanoislands to improve the efficiency of protein immobilization. By using HRP and hepatitis B virus surface antigen (HBsAg) as model analytes, the effect of the parylene-A film on the capacitive detection of protein adsorption was demonstrated.

Hydrogel-based diffusion chip with Electric Cell-substrate Impedance Sensing (ECIS) integration for cell viability assay and drug toxicity screening.

In this study, we have provided a novel analytical integration between hydrogel-based cell chip and Electric Cell-substrate Impedance Sensing (ECIS) technique to apply to a high-throughput, real-time cell viability assay and drug screening. For simulating the drug diffusion model, we have developed a hydrogel-based tissue-mimicking structure with microfluidic channel, without unwanted flow, to generate a gradient concentration with long-term stability. Along the gradient line, four individual micro-electrodes were installed to record the impedance signal changes, which result from the cell viability under drug effects. By watching for cellular impedance changes, we successfully estimated the cytotoxicity of the treatment corresponding to the various concentration values of stimuli, generated by the diffusion process along the channel. Reliable IC50 values and time-dose relationships were also achieved. With the feature of real-time monitoring capability, the advantages of non-invasion, label-free detection, time saving and simple manipulation, our integrative device has become a promising high throughput cell-based on-chip platform for cell viability assay and drug screening.

Silver nanoflower-reduced graphene oxide composite based micro-disk electrode for insulin detection in serum.

Sensitive and selective determination of protein biomarkers remains a significant challenge due to the existence of various biomarkers in human body at a low concentration level. Therefore, new technologies were incessantly steered to detect tiny biomarkers at a low concentration level, yet, it is difficult to develop reliable, stable and sensitive detection methods for disease diagnostics. Therefore, the present study demonstrates a methodology to detect insulin in serum at low levels based on Ag nanoflower (AgNF) decorated reduced graphene oxide (rGO) modified micro-disk electrode arrays (MDEAs). The morphology of AgNF-rGO composite was characterized by scanning electron microscopy, the structure was analyzed using X-ray diffraction patterns and Raman spectra. The hybrid interface exhibited enhanced electrical conductivity when compared with its individual elements and had improved capturing ability for antibody-antigen binding towards insulin detection. In order to measure quantitatively the insulin concentration in PBS and human serum, the change in impedance (ΔZ) from electrochemical impedance spectroscopy was analyzed for various concentrations of insulin in [Fe(CN)6](3-/4-) redox couple. The electrode with adsorbed antibodies showed an increase in ΔZ for the addition of antigen concentrations over a working range of 1-1000 ng mL(-1). The detection limits were 50 and 70 pg mL(-1) in PBS and human serum, respectively.

Label-free and direct detection of C-reactive protein using reduced graphene oxide-nanoparticle hybrid impedimetric sensor

For label-free and direct detection of C-reactive protein (CRP), an impedimetric sensor based on an indium tin oxide (ITO) electrode array functionalized with reduced graphene oxide-nanoparticle (rGO-NP) hybrid was fabricated and evaluated. Analytical measurements were performed to examine the properties of rGO-NP-modified ITO microelectrodes and to determine the influence upon sensory performance of using nanostructures modified for antibody immobilization and for recognition of CRP binding events. Impedimetric measurements in the presence of the redox couple [Fe(CN)6](3-/4-) showed significant changes in charge transfer resistance upon binding of CRP. The impedance measurements were highly target specific, linear with logarithmic CRP concentrations in PBS and human serum across a 1 ng mL(-1) and 1000 ng mL(-1) range and associated with a detection limits of 0.06 and 0.08 ng mL(-1) respectively.

Effect of cell senescence on the impedance measurement of adipose tissue-derived stem cells

Label-free and real-time monitoring of stem cells based on electrical impedance measurement is increasingly utilized for the quality control of the isolated stem cells to be used in stem cell-based tissue therapy or regenerative medicine. In spite of that the proliferative capacity and multipotency of stem cells are dependent on the type and age of the source tissue, however, the effect of the cell senescence on the impedance measurement of stem cells has not yet been studied. We investigated whether the senescence of adipose tissue-derived stem cells (ADSCs) can be detected by electrical impedance spectroscopy. For this, ADSCs at passage 9 and 31 were prepared and those genetic characteristics and growth kinetics were evaluated by quantitative polymerase chain reaction and cell counting. While the identified ADSCs were grown on the indium tin oxide electrodes, the impedance spectra were measured and interpreted by fitting analysis with an equivalent circuit model. ADSCs at passage 9 adhered on the electrode were small and spindle-shaped whereas the cells at passage 31 were flattened and larger than younger cells. At the beginning of culture time when the cell adhesion occurred, the resistance at 4.6kHz of passage 31 cells was higher than passage 9 due to the larger size of older cells. Afterwards, the value of passage 9 cells increased higher than passage 31, since younger cells proliferated more than old cells. Therefore, the impedance measurement could characterize the proliferative capacity of ADSCs during expanded culture.

Reduced Graphene Oxide Modified the Interdigitated Chain Electrode for an Insulin Sensor

Insulin is a key regulator in glucose homeostasis and its deficiency or alternations in the human body causes various types of diabetic disorders. In this paper, we present the development of a reduced graphene oxide (rGO) modified interdigitated chain electrode (ICE) for direct capacitive detection of insulin. The impedance properties of rGO-ICE were characterized by equivalent circuit modeling. After an electrochemical deposition of rGO on ICE, the electrode was modified with self-assembled monolayers and insulin antibodies in order to achieve insulin binding reactions. The impedance spectra and capacitances were measured with respect to the concentrations of insulin and the capacitance change (ΔC) was analyzed to quantify insulin concentration. The antibody immobilized electrode showed an increment of ΔC according to the insulin concentration in human serum ranging from 1 ng/mL to 10 µg/mL. The proposed sensor is feasible for label-free and real-time measuring of the biomarker and for point-of-care diagnosis.

Electrical impedance monitoring of C2C12 myoblast differentiation on an indium tin oxide electrode

Electrical cell-substrate impedance sensing is increasingly being used for label-free and real-time monitoring of changes in cell morphology and number during cell growth, drug screening, and differentiation. In this study, we evaluated the feasibility of using ECIS to monitor C2C12 myoblast differentiation using a fabricated indium tin oxide (ITO) electrode-based chip. C2C12 myoblast differentiation on the ITO electrode was validated based on decreases in the mRNA level of MyoD and increases in the mRNA levels of myogenin and myosin heavy chain (MHC). Additionally, MHC expression and morphological changes in myoblasts differentiated on the ITO electrode were comparable to those in cells in the control culture dish. From the monitoring the integration of the resistance change at 21.5 kHz, the cell differentiation was label-free and real-time detectable in 30 h of differentiation (p < 0.05).

Indium tin oxide based chip for optical and electrochemical characterization of protein–cell interaction

Analysis on the interaction between proteins and cells is required for understanding the cellular behaviour and response. In this article, we characterized the adhesion and growth of 293/GFP cells on fetal bovine serum (FBS) coated indium tin oxide (ITO) electrode. Using optical and electrochemical measurement, it was able to detect the adsorption of the protein on the surface of the ITO electrode dependent on the concentration of the protein in the immersing solution or the immersing time. An increase in the amount of the adsorbed serum protein resulted in a decrease in anodic peak current and an increase in the charge transfer resistance extracted from the equivalent circuit fitting analysis. More cells adhered and proliferated on the ITO electrode which was pre-immersed in FBS medium rather than bare electrode. The effect of the FBS on cell behaviors was reflected in the impedance monitoring of cells at 21.5 kHz.

Blood-based immunoassay of tau proteins for early diagnosis of Alzheimer's disease using surface plasmon resonance fiber sensors

We present the immunoassay of tau proteins (total tau and phosphorylated tau) in human sera using surface plasmon resonance (SPR) fiber sensors. This assay aimed at harvesting the advantages of using both SPR fiber sensors and a blood-based assay to demonstrate label-free point-of-care-testing (POCT) patient-friendly assay in a compact format for the early diagnosis of Alzheimers disease (AD). For conducting the assay, we used human sera of 40 subjects divided into halves, which were grouped into AD patients and control groups according to a number of neuropsychological tests. We found that on an average, the concentrations of both total tau and phosphorylated tau proteins (all known to be higher in cerebrospinal fluid (CSF) and the brain) turned out to be higher in human sera of AD patients than in controls. The limits of detection of total tau and phosphorylated tau proteins were 2.4 pg mL−1 and 1.6 pg mL−1, respectively. In particular, it was found that the AD group exhibited average concentration of total tau proteins 6-fold higher than the control group, while concentration of phosphorylated tau proteins was 3-fold higher than that of the control. We can attribute this inhomogeneity between both types of tau proteins (in terms of increase of control-to-AD in average concentration) to un-phosphorylated tau proteins being more likely to be produced in blood than phosphorylated tau proteins, which possibly is one of the potential key elements playing an important role in AD progress.

Recent Advances in Microfluidic Paper-based Electrochemiluminescence Analytical Devices for Point-of-care Testing Applications

Electrogenerated chemiluminescence (ECL) is an effective method for detecting a wide range of analytes including metal ions, virulent DNA, pathogenic bacteria, tumor cells and glucose. The attractive features of paper including passive liquid transport and biocompatibility are the main two advantages of using paper as a biosensing platform. To achieve key factors in paper-based sensors, the fabrication procedures and the analysis methods are fine tuned to satisfy the requirements of the ultimate-users. Here, we review various ECL signal amplification labels, inexpensive and portable devices, such as rechargeable batteries, which have replaced traditional instrumentation and different light detection technologies used in paper ECL devices. We also highlight the current trends and developments in ECL paper-based microfluidic analytical devices, as well as recent applications of ECL-based detection methods and inexpensive microfluidic devices. We discuss various paper-based devices, including 3D-origami devices, and devices utilizing self-powered and bipolar electrodes. Significant efforts have also been dedicated towards paper based multiplexing analysis (multi-label, and the multi-analyte strategies) and integration of microfluidic lab-on-paper devices with competences for point-to-care diagnostics. This review finally tabulates systematized data on figures of merit and novel types of ECL labels, used for detection of various biomarkers and analytes.