Imaging Biocatalytic Activity of Enzyme−Polymer Spots by Means of Combined Scanning Electrochemical Microscopy/Electrogenerated Chemiluminescence

Lei, Rong; Stratmann, Lutz; Schäfer, Dominik; Erichsen, Thomas; Neugebauer, Sebastian; Li, Na; Schuhmann, Wolfgang

doi:10.1021/ac900192n

Cited by 33 publications

(29 citation statements)

References 34 publications

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“…Measurements of local enzyme activity were similar to those obtained in SECM, with variations that were attributed to the different mechanisms involved in signal generation. Inhomogeneous activity was observed in the enzyme‐polymer spot, with a higher activity in the ring and a smaller one in the center (Figure ) …”

Section: Imaging Of Local Redox Activity/mapping Heterogeneitymentioning

confidence: 98%

Micro‐ and Nanoscopic Imaging of Enzymatic Electrodes: A Review

2019

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Redox enzymes, which catalyze electron transfer reactions in living organisms, can be used as selective and sensitive bioreceptors in biosensors, or as efficient catalysts in biofuel cells. In these bioelectrochemical devices, the enzymes are immobilized at a conductive surface, the electrode, with which they must be able to exchange electrons. Different physicochemical methods have been coupled to electrochemistry to characterize the enzyme‐modified electrochemical interface. In this Review, we summarize most efforts performed to investigate the enzymatic electrodes at the micro‐ and even nanoscale, thanks to microscopy techniques. Contrary to electrochemistry, which gives only a global information about all processes occurring at the electrode surface, microscopy offers a spatial resolution. Several techniques have been implemented; mostly scanning probe microscopies like atomic force microscopy, scanning tunneling microscopy, and scanning electrochemical microscopy, but also scanning electron microscopy and fluorescence microscopy. These studies demonstrate that various information can be obtained thanks to microscopy at different scales. Electrode imaging has been performed to confirm the presence of enzymes, to quantify and localize the biomolecules, but also to evaluate the morphology of immobilized enzymes, their possible conformation changes upon turnover, and their orientation at the electrode surface. Local redox activity has also been imaged and kinetics has been resolved.

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Section: Imaging Of Local Redox Activity/mapping Heterogeneitymentioning

confidence: 98%

Micro‐ and Nanoscopic Imaging of Enzymatic Electrodes: A Review

2019

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“…Since then, different active enzymes, including diaphorase [90,91], HRP [92,93], glucose dehydrogenase [94], ceruloplasmin [95], and cytochrome c peroxidase [96] have been investigated via SECM with different operating modes, and enzymatic sensing has become a major application for SECM in the biological field.…”

Section: Applicationsmentioning

confidence: 99%

Recent Advances in Scanning Electrochemical Microscopy for Biological Applications

Huang

Lou

et al. 2018

Materials

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Scanning electrochemical microscopy (SECM) is a chemical microscopy technique with high spatial resolution for imaging sample topography and mapping specific chemical species in liquid environments. With the development of smaller, more sensitive ultramicroelectrodes (UMEs) and more precise computer-controlled measurements, SECM has been widely used to study biological systems over the past three decades. Recent methodological breakthroughs have popularized SECM as a tool for investigating molecular-level chemical reactions. The most common applications include monitoring and analyzing the biological processes associated with enzymatic activity and DNA, and the physiological activity of living cells and other microorganisms. The present article first introduces the basic principles of SECM, followed by an updated review of the applications of SECM in biological studies on enzymes, DNA, proteins, and living cells. Particularly, the potential of SECM for investigating bacterial and biofilm activities is discussed.

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“…PMT biased at a high voltage is one of the most sensitive devices for light detection and capable of detecting single photon [3]. PMT can capture the ECL generated from localized surfaces, resulting in images of the substrate patterns [56][57][58] and visualizing the biocatalytic activity of enzyme-polymer spots [59]. CCD camera offers the advantages of instant image construction, high temporal and spatial resolution, and compatibility for highthroughput analysis.…”

Section: Ecl Imaging Analysismentioning

confidence: 99%

“…Sweat fingermarks contain much less chemicals, some of which, such as amino acids, can be tagged by ECL luminophores. Then the immobilized luminophores on ridge deposits react with free-diffusing co-reactants in solutions to emit light, generating a bright fingermark image against the dark background (designated as the positive imaging mode) [59,68]. Note that this approach works only directly for fingermarks found on conductive surfaces.…”

Section: Imaging Reactivity At Electrode Surfacesmentioning

confidence: 99%

Imaging Analysis Based on Electrogenerated Chemiluminescence

et al. 2017

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Electrochemiluminescence (ECL), also called electrogenerated chemiluminescence, is luminescence that is produced by chemical reactions triggered by electrochemical method. ECL imaging is a novel imaging technique, which can simultaneously provide two kinds of signals of both electrochemistry and optical image. Over the past two decades, ECL imaging has been not only a powerful tool for investigating the fundamental scientific questions such as ECL mechanisms and reaction kinetics, but also a versatile analytical technique for detection of a wide range of analytes including small molecules, DNA, proteins, and cells. In the first part of this review, we briefly describe the reaction mechanisms of ECL generation. Then the review focuses on the research progress on the ECL imaging approach. It is basically introduced from the following five aspects: (i) visualization of electroactive sites on surfaces, (ii) imaging analysis at the single-bead and single-cell levels, (iii) array bioanalysis, (iv) multi-color ECL imaging, and (v) paper chip based on ECL imaging. Finally, some perspectives and future directions in this active research area are presented.

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Imaging Biocatalytic Activity of Enzyme−Polymer Spots by Means of Combined Scanning Electrochemical Microscopy/Electrogenerated Chemiluminescence

Cited by 33 publications

References 34 publications

Micro‐ and Nanoscopic Imaging of Enzymatic Electrodes: A Review

Micro‐ and Nanoscopic Imaging of Enzymatic Electrodes: A Review

Recent Advances in Scanning Electrochemical Microscopy for Biological Applications

Imaging Analysis Based on Electrogenerated Chemiluminescence

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