Real-Time Monitoring of Potassium Ion Release Due to Apoptosis with Cell-Based Transparent-Gate Transistor

Sakata, Toshiya; Makino, Izumi; Sugimoto, Hitoshi

doi:10.1143/apex.5.017001

Cited by 9 publications

(9 citation statements)

References 13 publications

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“…In addition, ISFET sensors can be fabricated in large numbers using standard lithographic processes. Recent studies have demonstrated the use of ISFETs for measuring ion flux from single cells plated and grown on coated (Straub et al, 2001; Wrobel et al, 2005; Chang et al, 2012) or uncoated (Yu et al, 2007; Gebinoga et al, 2010; Sakata et al, 2012) sensors. However, the application of ISFETs in multi-well plate, parallel measurement configurations for ion efflux analysis has not been determined.…”

Section: Introductionmentioning

confidence: 99%

Application of ion-sensitive field effect transistors for ion channel screening

Walsh

DeRoller

Zhu

et al. 2014

Biosensors and Bioelectronics

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Cell-based screening assays are now widely used for identifying compounds that serve as ion channel modulators. However, instrumentation for the automated, real-time analysis of ion flux from clonal and primary cells is lacking. This study describes the initial development of an ion-sensitive field effect transistor (ISFET)-based screening assay for the acquisition of K+ efflux data from cells cultured in multi-well plates. Silicon-based K+-sensitive ISFETs were tested for their electrical response to varying concentrations of KCl and found to display a linear response relationship to KCl in the range of 10 µM to 1 mM. The ISFETs, along with reference electrodes, were inserted into fast-flow chambers containing either human colonic T84 epithelial cells or U251-MG glioma cells. Application of the Ca2+ ionophore A23187 (1 µM), to activate Ca2+-activated non-selective cation (NSC) channels (T84 cells) and large conductance Ca2+-activated K+ (BK) channels (U251 cells), resulted in time-dependent increases in the extracellular K+ concentration ([K+]o) as measured with the ISFETs. Treatment of the cells with blockers of either the NSC or BK channels, caused a strong inhibition of the A23187-induced increase in [K+]o. These results were consistent with ion current measurements obtained using the whole-cell arrangement of the patch clamp procedure. In addition, K+ efflux data could be acquired in parallel from multiple cell chambers using the ISFET sensors. Given the non-invasive properties of the probes, the ISFET-based assay should be adaptable for screening ion channels in various cell types.

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Section: Introductionmentioning

confidence: 99%

Application of ion-sensitive field effect transistors for ion channel screening

Walsh

DeRoller

Zhu

et al. 2014

Biosensors and Bioelectronics

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“…; Sakata & Miyahara ; Sakata et al . , ), based on the detection of changes in ionic charge density induced by cellular activities. In our previous works, field‐effect transistor (FET) biosensors monitored some cellular functions in real time, such as cellular respiration and programmed cell death (apoptosis), on the basis of ionic charges (Sakata & Miyahara ; Sakata et al .…”

Section: Introductionmentioning

confidence: 99%

“…In our previous works, field‐effect transistor (FET) biosensors monitored some cellular functions in real time, such as cellular respiration and programmed cell death (apoptosis), on the basis of ionic charges (Sakata & Miyahara ; Sakata et al . , ). Such ionic charges have to be selectively detected around a cell/sensor interface depending on the cellular functions of interest.…”

Section: Introductionmentioning

confidence: 99%

“…As an example of such biosensors, semiconductor-based biosensors enable the detection of cellular functions in a real-time, label-free and noninvasive manner (Fromherz et al 1991;Peitz et al 2007;Sakata & Miyahara 2008;Sakata et al 2012Sakata et al , 2013, based on the detection of changes in ionic charge density induced by cellular activities. In our previous works, field-effect transistor (FET) biosensors monitored some cellular functions in real time, such as cellular respiration and programmed cell death (apoptosis), on the basis of ionic charges (Sakata & Miyahara 2008;Sakata et al 2012Sakata et al , 2013. Such ionic charges have to be selectively detected around a cell/sensor interface depending on the cellular functions of interest.…”

Section: Introductionmentioning

confidence: 99%

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In situ electrical monitoring of cancer cells invading vascular endothelial cells with semiconductor‐based biosensor

Sakata

Matsuse

2017

Genes to Cells

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Cellular dynamics is very closely related to ionic behaviors, most of which have been hardly monitored in real time, whereas semiconductor-based biosensors have the unique advantage of direct detection of ionic charges in a real-time and noninvasive manner. In this study, we monitored the invasion process of cancer cells into the vascular endothelial layer in real time by a label-free method using a field-effect transistor (FET) biosensor. Endothelial cells were cultured on the sensing surface of the FET gate, to form a basement membrane between the endothelial cells and the sensing surface. When invasive cancer cells (HeLa cells) approached the endothelial cell-coated gate FET biosensor, a change in the surface potential was clearly detected using the FET biosensor. This is because HeLa cells, which invaded the endothelial cell layer, reduced the molecular charge density in the basement membrane by decomposing it. A platform based on the cell-coated gate FET biosensor is suitable for real-time and noninvasive monitoring of cellular dynamics based on intrinsic ionic charges.

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“…Field effect transistor (FET)-based biosensing systems have been proposed for some applications such as label-free DNA sequencing, [15][16][17] immunological assay [18] and cellular functional analysis. [19][20][21] In particular, FET-based biosensors are suitable for the detection of small molecules such as glucose as long as they have molecular charges. In the case of saccharide sensing, the coating of phenylboronic acid (PBA) derivatives on the gate surface is effective for the detection of low-molecular-weight saccharides such as glucose based on the principle of FETs, because the binding of PBA molecules with saccharide increases the density of negative charges in their conjugate on the basis of the equilibrium reaction.…”

Section: Introductionmentioning

confidence: 99%

Fundamental Characteristics of a Glucose Transistor with a Chemically Functional Interface

2017

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Semiconductor‐based biosensors are suitable for the detection of small molecules such as glucose as long as they have molecular charges. However, biological samples include various ionic molecules such as proteins; thus, their nonspecific adsorption should be prevented to reduce background noise. In this study, we optimized a field‐effect transistor (FET)‐based glucose sensor, that is, a glucose transistor, for more precise and highly sensitive glucose sensing. Using the glucose transistor modified with 4‐mercaptophenylboronic acid (PBA) on a gold gate electrode, we found that the detection sensitivity to glucose was increased to approximately 60 mV/decade from about 40 μM. This means that the PBA on the gate induced a diol‐binding site with glucose through an equilibrium reaction, resulting in an increase in the density of negative charges in the PBA‐glucose conjugate. Moreover, an oligoethyleneglycol (OEG) monolayer was coated on the gate as a molecular sieve interface so that the PBA molecules were embowered in the OEG membrane, but low‐molecular‐weight glucose could pass between the OEG molecules and reach the PBA molecules at the gate. The detection sensitivity of the biosensor with the optimized glucose transistor to glucose remained, to some extent, even in a solution including albumin. Thus, a platform based on the optimized glucose transistor is available for the practical use of a novel sensor that can detect glucose even at the low concentrations included in biological fluids.

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Real-Time Monitoring of Potassium Ion Release Due to Apoptosis with Cell-Based Transparent-Gate Transistor

Cited by 9 publications

References 13 publications

Application of ion-sensitive field effect transistors for ion channel screening

Application of ion-sensitive field effect transistors for ion channel screening

In situ electrical monitoring of cancer cells invading vascular endothelial cells with semiconductor‐based biosensor

Fundamental Characteristics of a Glucose Transistor with a Chemically Functional Interface

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