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Preliminary study to detect quantity of erythrocyte using interdigitated electrode by electrical impedance spectroscopy method
Abstract
Erythrocyte quantity is one of the parameters of the physiological conditions. This case causes knowledge about the nature and amount of erythrocytes to be interesting objects of research. This paper discusses the detection of erythrocyte quantities using an interdigitated electrode by Electrical Impedance Spectroscopy (EIS) method. Impedance measurements performed by injecting a current of 10 µA in the frequency range of 100 Hz - 100 kHz. Measurement made at erythrocyte concentrations from 3.79 x 10⁴cell µL-¹ to 7.61 × 10⁶ cell µL⁻¹. The results show that increasing the number of erythrocytes will increase the impedance value exponentially. Increasing the quantity of erythrocytes can be observed well at frequencies above 1 kHz. The minimum detection limit for erythrocytes is 3.79 x 10⁴cell µL⁻¹.
... In biological samples of whole blood (WB), the characteristic impedance can be affected by several factors: The number and morphology of red blood cells and blood plasma. Blood is a suspension of particles (red blood cells) with high resistivity in the conducting fluid (plasma) [20,21]. Other cells and platelets are considered to have an insignificant role in the electrical properties of blood because they are too small in size and number [22]. ...
... The frequency response test was carried out in the system calibrated to the AC source. Interdigitated electrodes were used, and the measurement mechanism was presented in previous studies [20,21]. The EIS Spectrum Analyzer application analyzes measurement results using an analogy model of an electrical circuit, as shown in Figure 1. ...
Doctors or health laboratory workers usually carry out manual blood examinations, who cell morphology parameters by eye. Electrical impedance spectroscopy has been proposed to assist doctors in examining blood damage. The effect of storage on blood cells will be evaluated using an electrical impedance measurement system on blood cells. The results of electrical impedance measurements will be correlated with the quality and quantity of blood cells, namely the mean corpuscular hemoglobin concentration (MCHC) value. Three bags of complete blood cell packages (from different donors) and 1 bag of red blood cell packages were used as blood samples. All blood samples using citrate phosphate dextrose adenine (CPDA) anticoagulant were obtained from the Indonesian Red Cross, Malang City Branch. Electrical impedance and cell-count measurements were performed on Day 0 (before being put into the refrigerator) and on Days 2, 4, 7, 10, 17, 21 and 28 after being placed in the fridge. Electrical impedance was measured using Bio-Impedance Spectroscopic Data Acquisition (BISDAQ). As the duration of storage increases, the MCHC value tends to decrease. The total impedance tends to decrease with increasing storage duration; the total impedance on Day 28 shows the lowest value. In general, the resistance value of the solution is correlated with the MCHC value of the blood cells. The escape of some material in the cell to the solution outside the cell results in a decrease in the resistance properties of the solution, according to the study’s results. The total electrical impedance decreases with increasing storage duration. Storage duration affects the quality and quantity of blood cells. The MCHC value of blood cells decreased with increasing storage duration. The value of MCHC is strongly correlated with the value of the external solution resistance of the cell. HIGHLIGHTS The total electrical impedance decreases with increasing storage duration Storage duration affects the quality and quantity of blood cells The MCHC value is strongly correlated with the value of the external solution resistance of the cell. The decrease in the MCHC value results in a reduction in the value of the external solution resistance of the cell GRAPHICAL ABSTRACT
Saccharin additives addition as a substitute for natural sweeteners is done to reduce production costs. This paper will discuss the effect of sweetener on the electric impedance of apple cider. The electrical impedance spectroscopy method was applied to the sample using four electrodes. Electrical impedance measurements made by injecting 100 μA currents in samples within the chamber in the frequency up to 1 MHz. The results showed that the electrical impedance value of apple cider manalagi is 1721.166 Ω to 41.870 Ω. The effect of sugar cane s mass composition and saccharin mass added to apple cider can be observed well in low frequency up to 10 kHz. The electric impedance characteristic of the apple cider manalagi increases with the addition of sugar canes mass, whereas the electric impedance value of the apple juice manalagi decreases with the addition of saccharin mass.
Electrical Impedance of biological tissues changes with the change in tissue anatomy and tissue physiology. Electrical Impedance Spectroscopy (EIS) has been studied to characterize the biological objects like fruits and vegetables for assessing their freshness. In this direction, the EIS studies have been conducted as a non-destructive investigation evaluation method to study the electrical impedance variations in banana ripening. The aim of the work is to correlate the impedance variation with the ripening process in banana. A small amount of alternating current is injected to the banana attached to an array of Ag/AgCl electrodes and the surface potentials are measured using Agilent 4294A impedance Analyzer. The banana impedance and phase angles are measured from 50 Hz and 1 MHz. The results demonstrate that the electrical impedance of banana varies significantly during its ripening. From the experimental studies, it is observed that the impedance, real part and imaginary part of the impedance all are increased with the progresses in the banana ripening process.
Impedance measurement of live biological cells is widely accepted as a label free, non-invasive and quantitative analytical method to assess cell status. This method is easy-to-use and flexible for device design and fabrication. In this review, three typical techniques for impedance measurement, i.e., electric cell-substrate impedance sensing, Impedance flow cytometry and electric impedance spectroscopy, are reviewed from the aspects of theory, to electrode design and fabrication, and applications. Benefiting from the integration of microelectronic and microfluidic techniques, impedance sensing methods have expanded their applications to nearly all aspects of biology, including living cell counting and analysis, cell biology research, cancer research, drug screening, and food and environmental safety monitoring. The integration with other techniques, the fabrication of devices for certain biological assays, and the development of point-of-need diagnosis devices is predicted to be future trend for impedance sensing techniques.
We illustrate a novel impedance immunosensor which rapidly and sensitively detects typhoid-causing infectious bacteria Salmonella enterica serovar (Salmonella typhi) in 10μL of sample volume. The bacteria are tagged with gold nanoparticles (Au NPs) via high-affinity antigen-antibody interactions for enhanced signal amplification and selectivity. The cell-particle bioconjugates are then subjected to alternating current (AC) electric fields applied through interdigitated microelectrodes. The immunosensor performance is optimized with respect to electric field frequency, cell concentration, incubation times and the type of blocking agent to achieve a low limit of detection (LOD) of 100CFU/mL. The approach is extendable to a wide spectrum of clinical diseases and offers an efficient and cost-effective solution for point-of-care diagnosis.
The objective of this work was to study the use of electrical impedance spectroscopy (EIS) measurement to characterize muscle electrical properties in different conditions.
In vivo EIS measurements were carried out in the range 1–60 kHz on 32 forearm flexor muscles of healthy volunteers. Each subject underwent to a protocol consisting of three trials: rest condition, isometric contraction, 4 min relaxation. Measured data were fitted with Cole–Cole equivalent electrical circuit using the complex nonlinear least square method.
Finally, the relative variations from the steady state of the electrical parameters of the equivalent electrical circuit were estimated.
Results obtained are in agreement with the hypothesis that, in the frequency range considered, the current preferentially flows through the extracellular fluid, being the component of intracellular resistance negligible. Among circuital parameters, in fact, the relative variations from steady state of R0 changed highly significant (p < 0.001) both during contraction and after 4 min relaxation.
Aims
Aplastic anaemia is a rare disorder characterized by peripheral pancytopenia and hypocellular bone marrow. Recent advancement of miRNA technologies, new promising therapy using small molecule inhibitors was suggested as efficient treatment option. Therefore, the study was undertaken to identify the significantly altered miRNA (miR-1202-upregulation) among aplastic anaemia patients compared to healthy controls by global miRNA expression profiling of bone marrow.
Materials and methods
miRNA and gene expression profiles for all the categories of aplastic anaemia patients and healthy controls were generated using Affymetrix probes.
Key findings
The study was based on freely available miRNA and host gene expression in NCBI GEO dataset. Microarray based gene expression profiling (GSE3807) revealed that RAPGEF5 and MANEA genes were significantly downregulated among aplastic anaemia patients compared to healthy controls and the expression of these genes were again upregulated after application of therapy among those patients compared to pre-therapy condition. RAPGEF5 was involved in Rap1 and Ras signaling pathways those were significantly enriched among aplastic anaemia patients and could be relevant for that phenotype. Microarray based miRNA expression profiling (GSE82095) also identified that miR-1202 was significantly upregulated among aplastic anaemia patients compared to controls and can potentially target RAPGEF5 and MANEA genes based on target prediction of miRNAs.
Significance
Thus synthetic miRNA inhibitors of miR-1202 can be used as a possible therapeutic agent to target miR-1202 and this inhibition can lead to its corresponding target gene upregulation for reversal of disease phenotype.
NaCl solution is a strong electrolyte that can produce Na⁺ and Cl⁻ ions. Measurements of impedance values of dielectric materials have been widely applied, but for materials containing many ions such as electrolyte solutions still carried out today. This paper analyzes the relationship between concentrations and the electrical impedance value of NaCl solutions. Electrochemical impedance spectroscopy (EIS) method is applied by injecting an electrochemical stimulus in the form of an electric current of one milli-ampere into NaCl solutions. Response output voltage was recorded using PicoScope S-5000. The impedance measurement uses two needle electrodes made of gold in the frequency range 1 Hz to 1 MHz. The sample of NaCl solutions used in NaCl and water ratio is 0.01-3.00%. The electrical impedance of the NaCl solutions decreases with the increase in concentration of NaCl solutions.
Traditional methods for detection of metabolically-active bacterial cells, while effective, require several days to complete. Development of sensitive electrical biosensors is highly desirable for rapid detection and counting of pathogens in food, water, or clinical samples. Herein, we develop a highly-sensitive non-Faradaic impedance sensor which detects metabolic activity of E. coli cells in a mere 1 μl of sample volume and without any sample filtration/purification. The three dimensional (3D) interdigitated electrodes (IDEs) along with self-assembled gold-nickel (Au-Ni) nanostructures significantly amplify the sensitivity by increasing the sensing area almost three-fold. The developed microsystem is integrated with an agar-based growth medium and monitors the metabolism of bacterial cells, enabling bacterial detection in approximately one hour after inoculation, i.e. in the lag-phase. Incorporation of a secondary agar layer as a biocompatible passivation layer protects the IDEs from potential Faradaic reactions and enhances sensitivity to modulation of the non-Faradaic impedance due to cellular metabolism. The resultant label-free sensor is capable of selective identification of metabolizing cells (vs. dead cells) across a wide linear range (10–1000 cells/μl). These results help pave the way for rapid antibacterial susceptibility testing at the point-of-need, which is currently a major challenge in healthcare.
In this study, distinguishing skin cancer cells (A431) and normal cells (HaCaT) was achieved using electrical impedance spectroscopy (EIS) with a novel developed device. The proliferation behaviors of the two cell types during a culture period of 5 days were characterized by the normalized impedance measured at 1465 Hz with simultaneous microscopic imaging for assistance. By fitting to the established equivalent circuits, A431 cells generated smaller resistance (Rc) values with smaller increasing variation, and comparable capacitance (Cc) values with similar decreasing variation compared with HaCaT cells. Moreover, Cc values were linearly correlated to the cell number. The results indicate that these two cell types can be distinguished with EIS based on the differences in the values and variation trends of Rc and Cc during the proliferation process in a real-time and label-free manner. Our work supplies a useful analytical approach for skin cancer cell research and may facilitate the early diagnosis of skin cancer.
A simple method based on electrochemical impedance spectroscopy using a gold electrode modified with folic acid-functionalized gold nanoparticles was used to detect folate receptor-rich HeLa cells with high sensitivity. The electrochemical impedance of the cytosensor was linear in two concentration ranges of HeLa cells, ranging from 6 × 100 to 1 × 103 and 1 × 103 to 1 × 105 cells mL−1. The detection limit for HeLa cells was 6 cells mL−1. This method could be significant for future applications in early stage cancer diagnosis.
The storage lesion of packed red blood cells (PRBCs) consists of biochemical changes associated with increased inflammatory mediators and decreased oxygen-carrying capacity. The effect of storage on the coagulation system is less well studied. The purpose of this study is to determine how PRBC storage time affects the activated coagulation time (ACT), using an in vitro model.
Type-matched PRBCs and fresh-frozen plasma were mixed, warmed to 37°C, and recalcified. The microsample ACT was repeatedly determined by 2 methods (ACT+ and ACT-LR) at 1 week, 3 weeks, and 5 weeks after date of donation. Hematocrit, pH, and ionized calcium were measured to assess how the model compared to physiologic circumstances. Statistical analysis was performed using the Student 2-tailed t-test with unequal variance.
Hematocrit, pH, and ionized calcium were successfully maintained within narrow ranges. The model was notably acidotic, consistent with circumstances of shock and massive transfusion. There was no significant difference in the ACT+ between week 1 and week 3 (P = .183), but there was a significant difference between week 3 and week 5 (P = .029) and between week 1 and week 5 (P = .007). The ACT-LR showed a significant difference between week 1 and week 3 (P = .001), but not between week 3 and week 5 (P = .286). There was again a strong difference between week 1 and week 5 results (P = .011).
In an isolated in vitro model, the storage lesion of PRBCs is associated with decreased coagulation. This may have relevance for transfusion practices in coagulation-sensitive circumstances such as trauma.
The possible influence of the cell shape on the derivation of the passive electrical parameters of a biological cell membrane is discussed in light of two different models which describe the cell as a shelled ellipsoidal particle and as a biconcave disk obtained by the revolution of the Cassini oval, respectively. Whereas within the first model, the Laplace equation can be solved analytically, in the second one a numerical algorithm based on the boundary element method has been employed. We have compared the results obtained by these two different models in the case of normal human erythrocyte cell membrane, using radiowave dielectric spectroscopy measurements. Our findings show that, although in principle the cell shape might deeply affect the evaluation of the passive electrical parameters of the cell membrane, in the case of the erythrocyte shape modelled by the Cassini curve, only small deviations are evidenced in comparison to the values derived, as usually done in the dielectric spectroscopy of biological cell suspensions, from an ellipsoidal model analysis. This result gives further support to the reliability of the data reported in the literature based on an ellipsoidal shape erythrocyte model.
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