Aflatoxins are highly toxic fungal secondary metabolites that often contaminate food and feed commodities. An electrochemical immunosensor for the determination of aflatoxin B1 (AFB1) was fabricated by immobilizing monoclonal AFB1 antibodies onto a screen-printed gold electrode that was modified with carbo-methyldextran by N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide cross-linking. An electrochemical interfacial modelling of biomolecular recognition was suggested and reasonably interpreted. Impedance technology was employed for the quantitative determination of AFB1. The limit of detection concentration of AFB1 for standard solutions and spiked pistachio samples was 0.5 ng/mL and 1 ng/mL, respectively. The immunosensor was able to successfully determine AFB1 concentrations in the range of 4.56–50.86 ng/mL in unknown pistachio samples. Comparative chromatographic analysis revealed that AFB1 concentrations that were higher than 345 ng/mL were not within the immunosensor’s upper limits of detection. Selectivity studies against Ochratoxin A and Aflatoxin M1 demonstrated that the proposed AFB1 immunosensor was able to differentiate between these other fungal mycotoxins. The novel electrochemical immunosensor approach has the potential for rapid sample screening in a portable, disposable format, thus contributing to the requirement for effective prevention and the control of aflatoxin B1 in pistachios.
Cancer cell lines are important tools for anticancer drug research and assessment. Impedance measurements can provide valuable information about cell viability in real time. This work presents the proof-of-concept development of a bioelectrical, impedance-based analysis technique applied to four adherent mammalian cancer cells lines immobilized in a three-dimensional (3D) calcium alginate hydrogel matrix, thus mimicking in vivo tissue conditions. Cells were treated with cytostatic agent5-fluoruracil (5-FU). The cell lines used in this study were SK-N-SH, HEK293, HeLa, and MCF-7. For each cell culture, three cell population densities were chosen (50,000, 100,000, and 200,000 cells/100 μL). The aim of this study was the extraction of mean impedance values at various frequencies for the assessment of the different behavior of various cancer cells when 5-FU was applied. For comparison purposes, impedance measurements were implemented on untreated immobilized cell lines. The results demonstrated not only the dependence of each cell line impedance value on the frequency, but also the relation of the impedance level to the cell population density for every individual cell line. By establishing a cell line-specific bioelectrical behavior, it is possible to obtain a unique fingerprint for each cancer cell line reaction to a selected anticancer agent.
Cell-based biosensors appear to be an attractive tool for the rapid, simple, and cheap monitoring of chemotherapy effects at a very early stage. In this study, electrochemical measurements using a four-point probe method were evaluated for suspensions of four cancer cell lines of different tissue origins: SK–N–SH, HeLa, MCF-7 and MDA-MB-231, all for two different population densities: 50 K and 100 K cells/500 μL. The anticancer agent doxorubicin was applied for each cell type in order to investigate whether the proposed technique was able to determine specific differences in cell responses before and after drug treatment. The proposed methodology can offer valuable insight into the frequency-dependent bioelectrical responses of various cellular systems using a low frequency range and without necessitating lengthy cell culture treatment. The further development of this biosensor assembly with the integration of specially designed cell/electronic interfaces can lead to novel diagnostic biosensors and therapeutic bioelectronics.
In this work, the assessment of the interactions of a bioactive substance applied to immobilized cells in either a two-dimensional (2D) or three-dimensional (3D) arrangement mimicking in vivo tissue conditions is presented. In particular, dopamine (DA) was selected as a stimulant for the implementation of an impedance analysis with a specific type of neural cells (murine neuroblastoma). The aim of this study was the extraction of calibration curves at various frequencies with different known dopamine concentrations for the description of the behavior of dopamine applied to 2D and 3D cell cultures. The results present the evaluation of the mean impedance value for each immobilization technique in each frequency. The differential responses showed the importance of the impedance when frequency is applied in both 2D and 3D immobilization cases. More specifically, in 2D immobilization matrix impedance shows higher values in comparison with the 3D cell culture. Additionally, in the 3D case, the impedance decreases with increasing concentration, while in the 2D case, an opposite behavior was observed.
This study presents a bioelectric cell-based biosensor for the monitoring of the pyrethroid pesticide cypermethrin, a voltage-gated sodium channel blocker, in tobacco samples. For this purpose, neuroblastoma cells were used as biorecognition elements. The potential interference by the tobacco major alkaloid nicotine on the detection of cypermethrin was also studied. In addition, fluorescence microscopy revealed a specific pattern of neuroblastoma cell calcium efflux (Ca2+) after treatment with nicotine or cypermethrin. Finally, actual field-derived tobacco extracts were used for assessing matrix effects on the biosensor’s performance. The biosensor could detect cypermethrin in concentrations up to 1.5 μg mL−1 without being influenced by the presence of nicotine and possibly other tobacco alkaloids. Though not selective for cypermethrin, the neuroblastoma-based biosensor system appears to be a promising alternative to laborious analysis methodologies for rapid, high throughput and cost-efficient screening of this pyrethroid in tobacco samples in the near future.
1 November 2017 in Journal of Physics: Conference Series
Electrochemical Impedance Spectroscopy (EIS) has become a technique that is frequently used for biological assays. Impedance is defined as a complex – valued generalization of resistance and varies depending on its use per application field. In health sciences, bioimpedance is widely used as non-invasive and low cost alternative in many medical areas that provides valuable information about health status. This work focuses on assessing the effects of a bioactive substance applied to immobilized cells. Dopamine was used as a stimulant in order to implement impedance analysis with a specific type of cells. Dopamine constitutes one of the most important catecholamine neurotransmitters in both the mammalian central and peripheral nervous systems. The main purpose is to extract calibration curves at different frequencies with known dopamine concentrations in order to describe the behavior of cells applied to dopamine using an impedance measurement device. For comparison purposes, non-immobilized cells were tested for the same dopamine concentrations.