Development of a label-free ATP image sensor for analyzing spatiotemporal patterns of ATP release from biological tissues

Doi, Hideo; Parajuli, Bijay; Horio, Tomoko; Shigetomi, Eiji; Shinozaki, Youichi; Noda, Toshihiko; Takahashi, Kazuhiro; Hattori, Toshiaki; Koizumi, Schuichi; Sawada, Kazuaki

doi:10.1016/j.snb.2021.129686

Cited by 10 publications

(10 citation statements)

References 37 publications

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“…The response times to reach 95 % of the saturation values were roughly estimated to be 10 s, 95 s, and 125 s for 1 µM, 10 µM, and 100 µM, respectively. Although the response times are still long at this stage, it was suggested that it is mainly limited by the diffusion of the molecules inside into the enzyme-immobilizing membrane as reported recently [48]. The molecules instantly reach the sensor surface with a sufficiently thin enzyme membrane, and thus, the response time would be reduced.…”

Section: H 2 O 2 Sensing Characteristicsmentioning

confidence: 88%

“…In a recent development, the sensor array with 256 × 256 pixels shrunk its pixel area down to 2 µm with the temporal resolution of 0.5 ms [45], showing the potential as a high spatio-temporal pH imaging technique. It was also demonstrated that the array applies to the detection of biomolecules including ACh [46] and ATP [47,48], based on acid generation by an enzymatic reaction. Although the CTT sensors showed the potential for the imaging of NTs, they suffer from low output signals in the application to living organisms including cells and tissues [48].…”

Section: Introductionmentioning

confidence: 99%

“…It was also demonstrated that the array applies to the detection of biomolecules including ACh [46] and ATP [47,48], based on acid generation by an enzymatic reaction. Although the CTT sensors showed the potential for the imaging of NTs, they suffer from low output signals in the application to living organisms including cells and tissues [48]. This comes from the fact that the sensor utilized pH change to capture the signal of NTs.…”

Section: Introductionmentioning

confidence: 99%

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Redox Sensor Array with 23.5-μm Resolution for Real-Time Imaging of Hydrogen Peroxide and Glutamate Based on Charge-Transfer-Type Potentiometric Sensor

Iwata

Okumura

et al. 2021

Sensors

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Towards clarifying the spatio-temporal neurotransmitter distribution, potentiometric redox sensor arrays with 23.5-µm resolution were fabricated. The sensor array based on a charge-transfer-type potentiometric sensor comprises 128×128 pixels with gold electrodes deposited on the surface of pixels. The sensor output corresponding to the interfacial potential of the electrode changed logarithmically with the mixture ratio of K3Fe(CN)6 and K4Fe(CN)6, where the redox sensitivity reached 49.9 mV/dec. By employing hydrogen peroxidase as an enzyme and ferrocene as an electron mediator, the sensing characteristics for hydrogen peroxide (H2O2) were investigated. The analyses of the sensing characteristics revealed that the sensitivity was about 44.7 mV/dec., comparable to the redox sensitivity, while the limit of detection (LOD) was achieved to be 1 µM. Furthermore, the oxidation state of the electron mediator can be the key to further lowering the LOD. Then, by immobilizing oxidizing enzyme for H2O2 and glutamate oxidase, glutamate (Glu) measurements were conducted. As a result, similar sensitivity and LOD to those of H2O2 were obtained. Finally, the real-time distribution of 1 µM Glu was visualized, demonstrating the feasibility of our device as a high-resolution bioimaging technique.

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Section: H 2 O 2 Sensing Characteristicsmentioning

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Redox Sensor Array with 23.5-μm Resolution for Real-Time Imaging of Hydrogen Peroxide and Glutamate Based on Charge-Transfer-Type Potentiometric Sensor

Iwata

Okumura

et al. 2021

Sensors

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“…Several types of electrode array devices are available for bioanalysis. 12 Large-scale integration (LSI)-based devices consisting of electrode arrays have been used for various bioanalysis applications, such as cellular characterization, 13 label-free imaging of adenosine 5′-triphosphate, 14 proton imaging of dynamics in the living brain, 15 tumor cell counting, 16 bacteria counting, 17 DNA detection, 18 glucose detection, 19 and imaging of metabolites in biofilms. 20 , 21 We previously developed an LSI-based amperometric/potentiometric device containing 20 × 20 sensors with a pitch of 250 μm.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Imaging of Endothelial Permeability Using a Large-Scale Integration-Based Device

et al. 2021

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It is important to clarify the transport of biomolecules and chemicals to tissues. Herein, we present an electrochemical imaging method for evaluating the endothelial permeability. In this method, the diffusion of electrochemical tracers, [Fe(CN) 6 ] 4– , through a monolayer of human umbilical vein endothelial cells (HUVECs) was monitored using a large-scale integration-based device containing 400 electrodes. In conventional tracer-based assays, tracers that diffuse through an HUVEC monolayer into another channel are detected. In contrast, the present method does not employ separated channels. In detail, a HUVEC monolayer is immersed in a solution containing [Fe(CN) 6 ] 4– on the device. As [Fe(CN) 6 ] 4– is oxidized and consumed at the packed electrodes, [Fe(CN) 6 ] 4– begins to diffuse through the monolayer from the bulk solution to the electrodes and the obtained currents depend on the endothelial permeability. As a proof-of-concept, the effects of histamine on the monolayer were monitored. Also, an HUVEC monolayer was cocultured with cancer spheroids, and the endothelial permeability was monitored to evaluate the metastasis of the cancer spheroids. Unlike conventional methods, the device can provide spatial information, allowing the interaction between the monolayer and the spheroids to be monitored. The developed method is a promising tool for organs-on-a-chip and drug screening in vitro.

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“…By immobilizing the target molecule-degrading enzyme on the ion-sensitive membrane of the sensor array, label-free real-time imaging of biomolecules, such as ATP [ 25 ] and acetylcholine (ACh) [ 26 ], is enabled. Recently, visualization of the spatiotemporal pattern of extracellular ATP has been demonstrated in biological tissues without any labeling [ 27 ]. However, the sensor output signal decreases due to the influence of pH-buffer components [ 28 , 29 ], which are generally used in in vitro environments, and the pH value in solution.…”

Section: Introductionmentioning

confidence: 99%

CMOS-Based Redox-Type Label-Free ATP Image Sensor for In Vitro Sensitive Imaging of Extracellular ATP

Doi

Horio

Choi

et al. 2021

Sensors

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Adenosine 5′-triphosphate (ATP) plays a crucial role as an extracellular signaling molecule in the central nervous system and is closely related to various nerve diseases. Therefore, label-free imaging of extracellular ATP dynamics and spatiotemporal analysis is crucial for understanding brain function. To decrease the limit of detection (LOD) of imaging extracellular ATP, we fabricated a redox-type label-free ATP image sensor by immobilizing glycerol-kinase (GK), L-α-glycerophosphate oxidase (LGOx), and horseradish peroxidase (HRP) enzymes in a polymer film on a gold electrode-modified potentiometric sensor array with a 37.3 µm-pitch. Hydrogen peroxide (H2O2) is generated through the enzymatic reactions from GK to LGOx in the presence of ATP and glycerol, and ATP can be detected as changes in its concentration using an electron mediator. Using this approach, the LOD for ATP was 2.8 µM with a sensitivity of 77 ± 3.8 mV/dec., under 10 mM working buffers at physiological pH, such as in in vitro experiments, and the LOD was great superior 100 times than that of the hydrogen ion detection-based image sensor. This redox-type ATP image sensor may be successfully applied for in vitro sensitive imaging of extracellular ATP dynamics in brain nerve tissue or cells.

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Development of a label-free ATP image sensor for analyzing spatiotemporal patterns of ATP release from biological tissues

Cited by 10 publications

References 37 publications

Redox Sensor Array with 23.5-μm Resolution for Real-Time Imaging of Hydrogen Peroxide and Glutamate Based on Charge-Transfer-Type Potentiometric Sensor

Redox Sensor Array with 23.5-μm Resolution for Real-Time Imaging of Hydrogen Peroxide and Glutamate Based on Charge-Transfer-Type Potentiometric Sensor

Electrochemical Imaging of Endothelial Permeability Using a Large-Scale Integration-Based Device

CMOS-Based Redox-Type Label-Free ATP Image Sensor for In Vitro Sensitive Imaging of Extracellular ATP

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