Large‐Scale Synthesis of Functionalized Nanowires to Construct Nanoelectrodes for Intracellular Sensing

Wu, Wentao; Jiang, Hong; Qi, Yuting; Fan, Wen‐Ting; Yan, Jing; Liu, Yanling; Huang, Wei‐Hua

doi:10.1002/ange.202106251

Cited by 14 publications

(2 citation statements)

References 63 publications

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“…10 µm protruding out of the sealed micropipette (Figure 1C). 17,44 To ensure that no extracellular signals could be detected after inserting sealed SiC@Pt NWEs inside cells, their whole surfaces were first entirely insulated with a thin SU8-2002 photoresist layer. Dipping the tip of these mounted SiC@Pt NWEs in acetone dissolved the photoresist and exposed a ca.…”

Section: Resultsmentioning

confidence: 99%

Homeostasis inside Single Activated Phagolysosomes: Quantitative and Selective Measurements of Submillisecond Dynamics of Reactive Oxygen and Nitrogen Species Production with a Nanoelectrochemical Sensor

Jiang

et al. 2022

J. Am. Chem. Soc.

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Reactive oxygen and nitrogen species (ROS/RNS) are generated by macrophages inside their phagolysosomes. This production is essential for phagocytosis of damaged cells and pathogens, i.e., protecting our organism and maintaining immune homeostasis. The ability to quantitatively and individually monitor the four primary ROS/RNS (ONOO -, H2O2, NO, and NO2 -) with sub-millisecond resolution is clearly warranted to elucidate the still unclear mechanisms of their rapid generation and to track their concentrations variations over time inside phagolysosomes, in particular, to document the origin of ROS/RNS homeostasis during phagocytosis. A novel nanowire electrode (NWE) has been specifically developed for this purpose. It consisted of wrapping a SiC NW with a mat of 3 nm platinum nanoparticles (Pt NPs) whose high electrocatalytic performances allow the characterization and individual measurements of each of the four primary ROS/RNS. This allowed for the first time a quantitative, selective and statistically robust determination of the individual amounts of ROS/RNS present in single dormant phagolysosomes. Additionally, the sub-millisecond resolution of the nanosensor allowed confirmation and measurement of the rapid ability of phagolysosomes to differentially mobilize their enzyme pools of NADPH oxidases (NOX) and inducible nitric oxide synthases (iNOS) to finely regulate their homeostasis. This reveals an essential key to immune responses and immunotherapies and rationalizes its biomolecular origin. ASSOCIATED CONTENT Supporting InformationThe Supporting Information is available free of charge at http://pubs.acs.org.Experimental details and supplement data including characterization analysis, electrochemical measurements and cell experiments (PDF)

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Section: Resultsmentioning

confidence: 99%

Homeostasis inside Single Activated Phagolysosomes: Quantitative and Selective Measurements of Submillisecond Dynamics of Reactive Oxygen and Nitrogen Species Production with a Nanoelectrochemical Sensor

Jiang

et al. 2022

J. Am. Chem. Soc.

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“…Meanwhile, the number of live-cell-permeant fluorophores that are available to perform intracellular sensing is still limited. Nanopipettebased electrochemical sensing provides an alternative to fluorescence-based assays by label-free and nondestructive measurement of the intracellular chemical reactivity of the target species [17]. Due to the small size of the tip, the damage to a single cell is minimal.…”

Section: Introductionmentioning

confidence: 99%

Robotic Intracellular Electrochemical Sensing for Adherent Cells

Zhan

et al. 2022

Cyborg Bionic Syst

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Nanopipette-based observation of intracellular biochemical processes is an important approach to revealing the intrinsic characteristics and heterogeneity of cells for better investigation of disease progression or early disease diagnosis. However, the manual operation needs a skilled operator and faces problems such as low throughput and poor reproducibility. This paper proposes an automated nanopipette-based microoperation system for cell detection, three-dimensional nonovershoot positioning of the nanopipette tip in proximity to the cell of interest, cell approaching and proximity detection between nanopipette tip and cell surface, and cell penetration and detection of the intracellular reactive oxygen species (ROS). A robust focus algorithm based on the number of cell contours was proposed for adherent cells, which have sharp peaks while retaining unimodality. The automated detection of adherent cells was evaluated on human umbilical cord vein endothelial cells (HUVEC) and NIH/3T3 cells, which provided an average of 95.65% true-positive rate (TPR) and 7.59% false-positive rate (FPR) for in-plane cell detection. The three-dimensional nonovershoot tip positioning of the nanopipette was achieved by template matching and evaluated under the interference of cells. Ion current feedback was employed for the proximity detection between the nanopipette tip and cell surface. Finally, cell penetration and electrochemical detection of ROS were demonstrated on human breast cancer cells and zebrafish embryo cells. This work provides a systematic approach for automated intracellular sensing for adherent cells, laying a solid foundation for high-throughput detection, diagnosis, and classification of different forms of biochemical reactions within single cells.

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Versatile Construction of Biomimetic Nanosensors for Electrochemical Monitoring of Intracellular Glutathione

Chen

Jiao

et al. 2022

Angewandte Chemie

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The current strategies for nanoelectrode functionalization usually involve sophisticated modification procedures, uncontrollable and unstable modifier assembly, as well as a limited variety of modifiers. To address this issue, we propose a versatile strategy for large-scale synthesis of biomimetic molecular catalysts (BMCs) modified nanowires (NWs) to construct functionalized electrochemical nanosensors. This design protocol employs an easy, controllable and stable assembly of diverse BMCs-poly(3,4-ethylenedioxythiophene) (PEDOT) composites on conductive NWs. The intrinsic catalytic activity of BMCs combined with outstanding electron transfer ability of conductive polymer enables the nanosensors to sensitively and selectively detect various biomolecules. Further application of sulfonated cobalt phthalocyanine functionalized nanosensors achieves real-time electrochemical monitoring of intracellular glutathione levels and its redox homeostasis in single living cells for the first time.

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Large‐Scale Synthesis of Functionalized Nanowires to Construct Nanoelectrodes for Intracellular Sensing

Cited by 14 publications

References 63 publications

Homeostasis inside Single Activated Phagolysosomes: Quantitative and Selective Measurements of Submillisecond Dynamics of Reactive Oxygen and Nitrogen Species Production with a Nanoelectrochemical Sensor

Homeostasis inside Single Activated Phagolysosomes: Quantitative and Selective Measurements of Submillisecond Dynamics of Reactive Oxygen and Nitrogen Species Production with a Nanoelectrochemical Sensor

Robotic Intracellular Electrochemical Sensing for Adherent Cells

Versatile Construction of Biomimetic Nanosensors for Electrochemical Monitoring of Intracellular Glutathione

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