New trends in enzyme-free electrochemical sensing of ROS/RNS. Application to live cell analysis

Rojas, Daniel; Hernández-Rodríguez, Juan F.; Pelle, Flavio Della; Escarpa, Alberto; Compagnone, Darío

doi:10.1007/s00604-022-05185-w

Cited by 9 publications

(4 citation statements)

References 112 publications

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“…After placing the cell culture dish on the micromanipulator, the nanoelectrode could be positioned at the subcellular level by adjusting the X/Y/Z axis with a remote control. 31,32 As shown in Figure 4A, the aptCFNE could easily penetrate into the nucleus, cytoplasm, and extracellular space (near the membrane) due to its conical and tiny tip. When using nanoelectrodes for intracellular studies, it was critical to ensure that the target cells were still alive after the insertion and electrochemical detection.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Assisted by a 3D micromanipulator, microscopy, and electrochemical workstation of the single-cell analyzer, this nanosensor was precisely moved to individual HeLa cells at the subcellular level, and electrochemical assays were carried out with good spatial resolution (Figure D,E). After placing the cell culture dish on the micromanipulator, the nanoelectrode could be positioned at the subcellular level by adjusting the X / Y / Z axis with a remote control. , As shown in Figure A, the aptCFNE could easily penetrate into the nucleus, cytoplasm, and extracellular space (near the membrane) due to its conical and tiny tip. When using nanoelectrodes for intracellular studies, it was critical to ensure that the target cells were still alive after the insertion and electrochemical detection.…”

Section: Resultsmentioning

confidence: 99%

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In Situ Measurement of ATP in Single Cells by an Amphiphilic Aptamer-Assisted Electrochemical Nano-Biosensor

Jiang

et al. 2022

Anal. Chem.

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In situ and quantitative measurements of adenosine 5′-triphosphate (ATP) in single living cells are highly desired for understanding several sorts of necessary physiological and pathological processes. Due to its small size and high sensitivity, an ultra-microelectrode can be used for single-cell analysis. However, ATP is difficult to detect in single cells because it is nonelectroactive and low in content. Herein, we introduced an electrochemical nano-biosensor based on an amphiphilic aptamer-assisted carbon fiber nanoelectrode (aptCFNE) with high signal-to-noise ratio. The low current (e.g., 60 pA) and the tiny diameter of the tip (ca. 400 nm) of the nanosensor made it noninvasive to living cells. The amphiphilic aptamer has good biocompatibility and can be stably modified to the surface of functionalized electrodes. CFNE, which was modified with ferrocene-labeled aptamer, could quickly and selectively detect ATP content in the nucleus, cytoplasm, and extracellular space of single HeLa cells. The results showed that the ATP contents in the nucleus, cytoplasm, and extracellular space were 568 ± 9, 461 ± 20, and 312 ± 4 μM, respectively. The anticancer drug treatment effects on the cellular level were further recorded, which was of great significance for understanding ATP-related biological processes and drug screenings. This strategy is universally applicable to detect other targets by changing the aptamer sequence, which will greatly improve our understanding of cell heterogeneity and provide a more reliable scientific basis for exploring major diseases at the single-cell level.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

In Situ Measurement of ATP in Single Cells by an Amphiphilic Aptamer-Assisted Electrochemical Nano-Biosensor

Jiang

et al. 2022

Anal. Chem.

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“…Various kinds of nanosystems based on metallic (metal organic frameworks, magnetic NPs, metal oxides) carbon (nanotubes, graphenes, Quantum dots), and biological (antibody, aptamer) nanomaterials are being gradually applied to electrochemical sensor and biosensors because of their incomparable physical, magnetic, chemical, and mechanical properties. The lifetime and the half-wave potential of the redox couple of biomarkers are crucial factors to detect oxidative stress biomarkers in biological conditions using electrochemical methods [18][19][20]. Activity retention of the labile biomolecules and reusability of a biosensor are two crucial parameters as the commercial viability of a device.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical approaches based on micro- and nanomaterials for diagnosing oxidative stress

et al. 2023

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This review article comprehensively discusses the various electrochemical approaches for measuring and detecting oxidative stress biomarkers and enzymes, particularly reactive oxygen/ nitrogen species, highly reactive chemical molecules, which are the byproducts of normal aerobic metabolism and can oxidize cellular components such as DNA, lipids, and proteins. First, we address the latest research on the electrochemical determination of reactive oxygen species generating enzymes, followed by detection of oxidative stress biomarkers, and final determination of total antioxidant activity (endogenous and exogenous). Most electrochemical sensing platforms exploited the unique properties of micro and nanomaterials such as carbon nanomaterials, metal or metal oxide nanoparticles (NPs), conductive polymers and metal-nano compounds, which have been mainly used for enhancing the electrocatalytic response of sensors/biosensors. The performance of the electroanalytical device commonly measured by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in terms of detection limit, sensitivity, and linear range of detection was also discussed. This article provides a comprehensive review of electrode fabrication, characterization and evaluation of their performances, which are assisting to design and manufacture an appropriate electrochemical (bio)sensor for medical and clinical applications. The key points such as accessibility, affordability, rapidity, low cost, and high sensitivity of the electrochemical sensing devices were also highlighted for the diagnosis of oxidative stress. Overall, this review brings a timely discussion on past and current approaches for developing electrochemical sensors and biosensors mainly based on micro and nanomaterials for the diagnosis of oxidative stress.

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“…This makes it very challenging to continuously monitor oxidative stress in real-time during cell growth [ 24 ]. Electrochemical sensors are perfect candidates to minimize all these drawbacks because they can be applied for in situ and real-time analysis, while offering good performances in terms of sensitivity, selectivity and limit of detection [ 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 ]. To improve the performance of an electrochemical cell, the use of nanostructured electrodes ensures a high active surface area and promotes a high current density [ 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 ].…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Quantification of H2O2 Released by Airway Cells Growing in Different Culture Media

et al. 2022

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Quantification of oxidative stress is a challenging task that can help in monitoring chronic inflammatory respiratory airway diseases. Different studies can be found in the literature regarding the development of electrochemical sensors for H2O2 in cell culture medium to quantify oxidative stress. However, there are very limited data regarding the impact of the cell culture medium on the electrochemical quantification of H2O2. In this work, we studied the effect of different media (RPMI, MEM, DMEM, Ham’s F12 and BEGM/DMEM) on the electrochemical quantification of H2O2. The used electrode is based on reduced graphene oxide (rGO) and gold nanoparticles (AuNPs) and was obtained by co-electrodeposition. To reduce the electrode fouling by the medium, the effect of dilution was investigated using diluted (50% v/v in PBS) and undiluted media. With the same aim, two electrochemical techniques were employed, chronoamperometry (CH) and linear scan voltammetry (LSV). The influence of different interfering species and the effect of the operating temperature of 37 °C were also studied in order to simulate the operation of the sensor in the culture plate. The LSV technique made the sensor adaptable to undiluted media because the test time is short, compared with the CH technique, reducing the electrode fouling. The long-term stability of the sensors was also evaluated by testing different storage conditions. By storing the electrode at 4 °C, the sensor performance was not reduced for up to 21 days. The sensors were validated measuring H2O2 released by two different human bronchial epithelial cell lines (A549, 16HBE) and human primary bronchial epithelial cells (PBEC) grown in RPMI, MEM and BEGM/DMEM media. To confirm the results obtained with the sensor, the release of reactive oxygen species was also evaluated with a standard flow cytometry technique. The results obtained with the two techniques were very similar. Thus, the LSV technique permits using the proposed sensor for an effective oxidative stress quantification in different culture media and without dilution.

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New trends in enzyme-free electrochemical sensing of ROS/RNS. Application to live cell analysis

Cited by 9 publications

References 112 publications

In Situ Measurement of ATP in Single Cells by an Amphiphilic Aptamer-Assisted Electrochemical Nano-Biosensor

In Situ Measurement of ATP in Single Cells by an Amphiphilic Aptamer-Assisted Electrochemical Nano-Biosensor

Electrochemical approaches based on micro- and nanomaterials for diagnosing oxidative stress

Electrochemical Quantification of H2O2 Released by Airway Cells Growing in Different Culture Media

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