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Cited by 22Electrochemical sensors and biosensorsElectrochemical Analysis and ApplicationsAdvanced biosensing and bioanalysis techniques

Trends in single-impact electrochemistry for bacteria analysis

Open access
Smida, Hassiba, Langlard, Arthur, Ameline, Dorine, Thobie-Gautier, Christine, Boujtita, Mohammed, Lebègue, Estelle·2023·Analytical and Bioanalytical Chemistry
DOI 10.1007/s00216-023-04568-z
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Abstract

Single-impact electrochemistry for the analysis of bacteria is a powerful technique for biosensing applications at the single-cell scale. The sensitivity of this electro-analytical method has been widely demonstrated based on chronoamperometric measurements at an ultramicroelectrode polarized at the appropriate potential of redox species in solution. Furthermore, the most recent studies display a continuous improvement in the ability of this sensitive electrochemical method to identify different bacterial strains with better selectivity. To achieve this, several strategies, such as the presence of a redox mediator, have been investigated for detecting and identifying the bacterial cell through its own electrochemical behavior. Both the blocking electrochemical impacts method and electrochemical collisions of single bacteria with a redox mediator are reported in this review and discussed through relevant examples. An original sensing strategy for virulence factors originating from pathogenic bacteria is also presented, based on a recent proof of concept dealing with redox liposome single-impact electrochemistry. The limitations, applications, perspectives, and challenges of single-impact electrochemistry for bacteria analysis are briefly discussed, based on the most significant published data.

AI snapshot

Written from the paper’s full text.

TL;DR

This review summarizes single-impact electrochemistry for bacteria analysis, covering blocking collisions, redox-mediator collisions, and redox-liposome detection of virulence factors. It reports high sensitivity at the single-cell scale but concludes that selective identification of a given bacterial strain in complex mixtures remains the main challenge.

Key findings

  • Blocking electrochemical impacts detect single bacteria as current steps when cells adsorb onto a polarized ultramicroelectrode (UME); Ronspees et al. observed step-shaped transients for *E. coli* but spike/blip-shaped transients for *B. subtilis* under the same FcM/10 μm Pt UME conditions, attributed to adsorption versus short-lived bouncing.
  • Redox-mediator collisions can probe bacterial electrochemical activity: Gao et al. detected *E. coli* via ferricyanide reduction and *S. maltophilia* via regeneration of Ru(NH₃)₆³⁺; Couto et al. used TMPD to detect *E. coli* cytochrome c oxidases and discriminate living from dead bacteria; Chen et al. quantified thionine reduction at 6.8×10⁻¹⁸ mol per *E. coli* versus 3.5×10⁻¹⁸ mol per *B. subtilis*.
  • A three-mediator system (K₃Fe(CN)₆, K₄Fe(CN)₆, and menadione) produced opposite collision signals for *S. cerevisiae* versus *E. coli* and *S. aureus*, enabling selective detection of *S. cerevisiae* in a mixed microbial suspension.
  • Redox DMPC liposome single-impact electrochemistry detected 500 nM rhamnolipid virulence factor in aqueous phosphate buffer in less than 30 minutes at a Pt UME.

Methods

  • Review article synthesizing the most significant published data on single-impact electrochemistry for bacteria analysis, focused on three strategies: blocking electrochemical impacts of single bacteria, electrochemical collisions with a redox mediator, and virulence-factor detection via redox liposome single-impact electrochemistry.
  • The reviewed measurements use chronoamperometry at polarized UMEs in aqueous electrolyte containing redox species and bacteria, typically at 10⁶ to 10⁹ bacteria per milliliter; examples include *E. coli*, *B. subtilis*, *S. oneidensis*, *L. lactis*, *S. maltophilia*, *S. cerevisiae*, *S. aureus*, and *P. aeruginosa* rhamnolipid.
  • Main analysis is qualitative and comparative, discussing representative studies, collision frequency–concentration relationships, current transient shapes, and performance metrics rather than presenting new primary experiments.

Limitations

  • Stated: the blocking collision signal does not allow identification of the adsorbed bacterium and is only suitable for discriminating single cells by electrostatic attraction/repulsion according to their charge and UME polarization.
  • Stated: single-impact electrochemistry has not proved efficient for selective detection yet, except for discrimination between Gram-positive and Gram-negative bacteria; redox-mediator methods can lack selectivity, and the three-mediator system may not suit similar bacterial strains.
  • Stated and evident: the virulence-factor liposome proof of concept needs extension to other toxins, liposome types, and complex supernatants; the review itself is not a primary study, and many conclusions rest on proof-of-concept experiments with model bacteria in simple aqueous solutions rather than real complex matrices.

Model-written summary. Check it against the paper before relying on it.

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Registry

Crossref · 10.1007/s00216-023-04568-z

Cites

44 of 44 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

23 of 79 cited sources checked · 16 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    The blocking electrochemical impacts method has been widely extended to the detection of various micro- and nano-objects, with specific interest in single collisions of bio-entities such as proteins, viruses, and bacteria.

    “In the following years, this method has been widely extended to the detection of various micro-and nano-objects, with a specific interest in single collisions of bio-entities such as proteins, viruses, and bacteria [19,25,29,35].”
    Quote in the paperNot a substantial claim3 of 4 sources checked
    • cites [19] Dick JE, Hilterbrand AT, Boika A, Upton JW, Bard AJ. Electrochemical detection of a single cytomegalovirus at an ultramicroelectrode and its antibody anchoring. Proc Natl Acad Sci. 2015;112(17):5303–8. source
    • cites [25] Dick JE, Renault C, Bard AJ. Observation of single-protein and DNA macromolecule collisions on ultramicroelectrodes. J Am Chem Soc. 2015;137(26):8376–9. source
    • cites [29] Lee JY, Kim BK, Kang M, Park JH. Label-free detection of single living bacteria via electrochemical collision event. Sci Rep. 2016;6:30022. source
    • cites [35] Deng Z, Renault C. Detection of individual insulating entities by electrochemical blocking. Curr Opin Electrochem. 2021;25: 100619. source
  • Premisecitation

    An original and recent approach is to detect virulence factors released by pathogenic bacteria rather than the cells themselves, based on redox liposomes single-impact electrochemistry.

    “Also, an original and recent approach is to detect the virulence factors released by pathogenic bacteria rather than the cells themselves, based on redox liposomes single-impact electrochemistry [17].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [17] Luy J, Ameline D, Thobie-Gautier C, Boujtita M, Lebègue E. Detection of bacterial rhamnolipid toxin by redox liposome single impact electrochemistry. Angew Chem Int Ed. 2022;61(6): e202111416. source
  • Premisecitation

    For single S. oneidensis impacts at the potential of steady-state current of potassium ferrocyanide (E ≥ +0.3 V vs Ag/AgCl), a typical stair-step current response is detected, corresponding to adsorption of cells onto the UME.

    “In this study, a typical stair-step current response is detected for single S. oneidensis impacts at the potential of steady-state current of potassium ferrocyanide (E ≥ +0.3 V vs. Ag/AgCl), corresponding to the adsorption of cells onto the UME [16].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [16] Lebègue E, Costa NL, Louro RO, Barrière F. Communication—electrochemical single nano-impacts of electroactive Shewanella oneidensis bacteria onto carbon ultramicroelectrode. J Electrochem Soc. 2020;167(10): 105501. source
  • Premisecitation

    Lee et al. demonstrated a linear relationship between collision frequency f_exp and E. coli concentration C_bac, as expected by Equation 1.

    “The authors have demonstrated a linear relationship between the collision frequency f exp (number of step-shaped transients) and the E. coli concentration C bac , as expected by Equation 1 below (in conditions where the mass transport is controlled by diffusion) [10,16,32].”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [10] Goines S, Dick JE. Review—electrochemistry’s potential to reach the ultimate sensitivity in measurement science. J Electrochem Soc. 2019;167(3): 037505. source

      In “Methods to Decrease Limit of Quantitation” (page unavailable), the review gives a diffusion-controlled equation in which collision frequency is proportional to the concentration of colliding entities. Its discussion of bacterial blocking experiments (page unavailable) reports detection of single E. coli collisions, but does not report that Lee et al. experimentally demonstrated a linear relationship between step-shaped transient frequency and E. coli concentration. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [16] Lebègue E, Costa NL, Louro RO, Barrière F. Communication—electrochemical single nano-impacts of electroactive Shewanella oneidensis bacteria onto carbon ultramicroelectrode. J Electrochem Soc. 2020;167(10): 105501. source

      The Summary uses a diffusion-based equation to relate the frequency of step-shaped impacts to bacterial concentration. But this paper measures Shewanella oneidensis at one reported concentration; it does not demonstrate an experimental linear relationship across E. coli concentrations. Pages were not provided in our copy. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [32] Couto RAS, Chen L, Kuss S, Compton RG. Detection of Escherichia coli bacteria by impact electrochemistry. Analyst. 2018;143(20):4840–3. source
  • Premisecitation

    The magnitude of current steps in blocking impact experiments depends on the size of the blocking object, the size of the UME, the type and concentration of the redox probe, and the location of the blocking object adsorbed onto the UME.

    “The magnitude of the current steps in blocking impact experiments depends on the size of the blocking object, the size of the UME, the type and the concentration of the redox probe, and the location of the blocking object adsorbed onto the UME [30,35].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [30] Ronspees AT, Thorgaard SN. Blocking electrochemical collisions of single E. coli and B. subtilis bacteria at ultramicroelectrodes elucidated using simultaneous fluorescence microscopy. Electrochim Acta. 2018;278:412‑20. source
    • cites [35] Deng Z, Renault C. Detection of individual insulating entities by electrochemical blocking. Curr Opin Electrochem. 2021;25: 100619. source

      In “Measuring the size of a particle by electrochemical blocking, an analytical challenge” (page unavailable), the paper links current-step magnitude to particle size, electrode size, redox-reporter concentration, and the particle’s position on the electrode. It does not establish that the type of redox probe affects step magnitude. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
  • May combine two claims. 15
  • Classified as probably not a substantial claim. 4

  • May combine two claims.

    Premisecitation

    Single-impact electrochemistry provides a low limit of detection (in principle, one single species) and the ability to study single entities in real-time through a dynamic measurement.

    “Single-impact electrochemistry provides a low limit of detection (in principle, one single species) inherent to this electro-analytical method and the ability to study single entities (cells, viruses, bacteria, nanoparticles…) in real-time through a dynamic measurement [12][13][14][15][16][17][18][19][20].”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites [12] Sekretareva A. Single-entity electrochemistry of collision in sensing applications. Sens Actuators Rep. 2021;3: 100037. source
    • cites [13] Lebègue E, Anderson CM, Dick JE, Webb LJ, Bard AJ. Electrochemical detection of single phospholipid vesicle collisions at a Pt ultramicroelectrode. Langmuir. 2015;31(42):11734–9. source
    • cites [14] Dick JE, Lebègue E, Strawsine LM, Bard AJ. Millisecond coulometry via zeptoliter droplet collisions on an ultramicroelectrode. Electroanalysis. 2016;28(10):2320–6. source

  • May combine two claims.

    Premisecitation

    Virulence factors such as RL are known to interact with the outer lipid bilayer membrane of eukaryotic cells, and this property can be extended to liposomes and vesicles for pathogenic bacteria sensing.

    “The virulence factors such as RL are known to interact with the outer lipid bilayer membrane of eukaryotic cells and this property can be extended to liposomes and vesicles for pathogenic bacteria sensing [39].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [39] Thet NT, Jenkins ATA. An electrochemical sensor concept for the detection of bacterial virulence factors from Staphylococcus aureus and Pseudomonas aeruginosa. Electrochem Commun. 2015;59:104–8. source

      The Abstract describes biomimetic lipid vesicles that release their contents in response to toxins, and reports testing with rhamnolipid and bacterial supernatants. It supports the vesicle-based sensing concept, but our copy does not state that rhamnolipid interacts with the outer lipid bilayer of eukaryotic cells. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Redox liposomes single-impact electrochemistry detects oxidation of ferrocyanide encapsulated in liposomes during single collisions onto a polarized UME surface, producing current spikes in the i-t curve.

    “Briefly, the redox liposomes singleimpact electrochemistry consists in detecting the oxidation of ferrocyanide encapsulated in liposomes during single collisions onto the polarized UME surface, corresponding to current spikes in the i-t curve [20].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [20] Smida H, Thobie-Gautier C, Boujtita M, Lebègue E. Recent advances in single liposome electrochemistry. Curr Opin Electrochem. 2022;36: 101141. source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    RL is a glycolipidic biosurfactant secreted by Pseudomonas aeruginosa, a pathogenic Gram-negative bacterium involved in nosocomial infections.

    “RL is a glycolipidic biosurfactant secreted by Pseudomonas aeruginosa (P.aeruginosa), pathogenic Gram-negative bacteria involved in nosocomial infections [17].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [17] Luy J, Ameline D, Thobie-Gautier C, Boujtita M, Lebègue E. Detection of bacterial rhamnolipid toxin by redox liposome single impact electrochemistry. Angew Chem Int Ed. 2022;61(6): e202111416. source

      The Abstract states that Pseudomonas aeruginosa secretes the glycolipidic biosurfactant rhamnolipid and is involved in nosocomial infections. It also describes the bacterium as pathogenic, but our copy does not identify it as Gram-negative; that part of the attributed claim is unsupported. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Blip-shaped collision transients for B. subtilis are due to short-lived bounce events of bacteria at the UME, in contrast with step-shaped transients due to adsorption of E. coli at the UME.

    “Based on the simultaneous fluorescence microscopy imaging (Fig. 1c), these blip-shaped collision transients are due to short-lived bounce events of bacteria at the UME, in contrast with the step-shaped transients observed in Fig. 1b due to the adsorption of E. coli at the UME [30].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [30] Ronspees AT, Thorgaard SN. Blocking electrochemical collisions of single E. coli and B. subtilis bacteria at ultramicroelectrodes elucidated using simultaneous fluorescence microscopy. Electrochim Acta. 2018;278:412‑20. source

  • May combine two claims.

    Interpretationcitation

    Ronspees et al. work provides the first example of different species of bacteria leading to different transients in a blocking collision experiment, and thus opens the way to selective detection of bacteria.

    “This work provides the first example of different species of bacteria leading to different transients (current signals) in a blocking collision experiment, and thus opens the way to the selective detection of bacteria [30].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [30] Ronspees AT, Thorgaard SN. Blocking electrochemical collisions of single E. coli and B. subtilis bacteria at ultramicroelectrodes elucidated using simultaneous fluorescence microscopy. Electrochim Acta. 2018;278:412‑20. source

  • May combine two claims.

    Premisecitation

    When bacteria enter the diffusion layer, a feedback redox cycle is created and a reductive current spike is detected, corresponding to regeneration of TMPD•+ in the periphery of the bacterium.

    “When the bacteria enter the diffusion layer, a feedback redox cycle is created and a reductive current spike (current increase) is detected in the chronoamperometry measurement, corresponding to the regeneration of TMPD •+ in the periphery of the bacterium (Fig. 3a) [32].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [32] Couto RAS, Chen L, Kuss S, Compton RG. Detection of Escherichia coli bacteria by impact electrochemistry. Analyst. 2018;143(20):4840–3. source

  • Classified as probably not a substantial claim. May combine two claims.

    Interpretationcitation

    This work demonstrates an interesting method based on the electrochemical collision technique for quantifying bacterial redox activity with a mediator at the single-cell level, which is of great significance to better understand electron transfer processes in microorganisms and thus improve the design of microbial electrochemical technologies.

    “This work demonstrates an interesting method based on the electrochemical collision technique for quantifying the bacterial redox activity with a mediator at the single-cell level, which is of great significance to better understand electron transfer processes in microorganisms and thus, improve the design of microbial electrochemical technologies [33].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [33] Chen Y, Wang D, Liu Y, Gao G, Zhi J. Redox activity of single bacteria revealed by electrochemical collision technique. Biosens Bioelectron. 2020;176: 112914. source

  • May combine two claims.

    Premisecitation

    Redox reactions between S. cerevisiae cells and mediators reduce K3Fe(CN)6 to K4Fe(CN)6 and increase the local concentration of K4Fe(CN)6.

    “However, the redox reactions between the S. cerevisiae cells and mediators also reduce K 3 Fe(CN) 6 to K 4 Fe(CN) 6 and increase the local concentration of K 4 Fe(CN) 6 at the same time.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [34] Chen Y, Liu Y, Wang D, Gao G, Zhi J. Three-mediator enhanced collisions on an ultramicroelectrode for selective identification of single saccharomyces cerevisiae. Anal Chem. 2022;94(37):12630–7. source

  • May combine two claims.

    Premisecitation

    Thionine can be reduced by the bacterial cell and when a single bacterium-TH complex collides onto the UME (5 μm carbon fiber), the reduced thionine is re-oxidized at +1.0 V vs Ag/AgCl and generates a current spike in the i-t curve.

    “Thionine can be reduced by the bacterial cell and when a single bacterium-TH complex collides onto the UME (5 µm diameter carbon fiber), the reduced thionine is re-oxidized at an appropriate potential (+1.0 V vs. Ag/AgCl) and generate a current spike in the i-t curve [33].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [33] Chen Y, Wang D, Liu Y, Gao G, Zhi J. Redox activity of single bacteria revealed by electrochemical collision technique. Biosens Bioelectron. 2020;176: 112914. source

  • May combine two claims.

    Premisecitation

    In the three-mediator system, E. coli and S. aureus only generate downward current steps because blocking of mediator diffusion suppresses their redox activities, so only blocking collisions are observed in their i-t curves.

    “Indeed, both E. coli and S. aureus only generate downward current steps because the blocking effect of mediator diffusion suppresses their redox activities (illustrated in Fig. 3b, bottom left for E. coli), thus only blocking collisions are observed in the i-t curves (shown in Fig. 3b, bottom right for E. coli) for these two types of bacteria in the three-mediator system [34].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [34] Chen Y, Liu Y, Wang D, Gao G, Zhi J. Three-mediator enhanced collisions on an ultramicroelectrode for selective identification of single saccharomyces cerevisiae. Anal Chem. 2022;94(37):12630–7. source

  • May combine two claims.

    Interpretationnone given

    Decorating E. coli cells with Ag nanoparticles requires pretreatment of the cells and is not easily applicable to selective bacteria detection and identification.

    “Nevertheless, this technique requires a pretreatment of the cells and is not easily applicable to selective bacteria detection and identification.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Menadione, a hydrophobic mediator, can selectively penetrate the S. cerevisiae membrane, access its intracellular redox center, and react with K3Fe(CN)6 in bulk solution.

    “The hydrophobic mediator (menadione) can selectively penetrate through the S. cerevisiae membrane and get access to its intracellular redox center, then can further react with K 3 Fe(CN) 6 in the bulk solution (Fig. 3b, top left) [34].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [34] Chen Y, Liu Y, Wang D, Gao G, Zhi J. Three-mediator enhanced collisions on an ultramicroelectrode for selective identification of single saccharomyces cerevisiae. Anal Chem. 2022;94(37):12630–7. source

  • May combine two claims.

    Premisecitation

    Fe(III) is reduced by E. coli bacteria colliding with the UME surface and the produced Fe(II) species get oxidized back at the UME.

    “In this case, Fe(III) is reduced by the E.coli bacteria colliding with the UME surface and the produced Fe(II) species get oxidized back at the UME (Fig. 2a) [28].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [28] Gao G, Wang D, Brocenschi R, Zhi J, Mirkin MV. Toward the detection and identification of single bacteria by electrochemical collision technique. Anal Chem. 2018;90(20):12123–30. source

  • May combine two claims.

    Interpretationreasoning

    The collision signal (current step transient) in blocking electrochemical impacts does not allow identification of the adsorbed bacterium, so the technique is only suitable for discriminating single cells by electrostatic attraction/repulsion according to their own charge and UME polarization.

    “Nevertheless, the collision signal (current step transient) does not allow to identify the adsorbed bacterium, thus this technique is only suitable for discriminating single cells by electrostatic attraction/repulsion according to their own charge and the UME polarization.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S4 (based).

    Interpretationnone given

    Single-impact electrochemistry is currently the most sensitive method to detect bacteria at the single-cell scale in a short analysis time.

    “This is currently the most sensitive method to detect bacteria at the single-cell scale in a short analysis time, however the selectivity still needs to be improved for a possible extension to sensor applications and fundamental purposes.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationnone given

    The two main current techniques in single-impact electrochemistry for bacteria analysis are blocking electrochemical impacts of single bacteria and electrochemical collisions with a redox mediator.

    “The two main current techniques in this area are the blocking electrochemical impacts of single bacteria and the electrochemical collisions with a redox mediator.”
    Quote in the paperSubstantial claim
17 of 17

  • The paper relies on this without support.

    Electron transfer processes for fungi and bacteria differ significantly.

    “However, the electron transfer processes for fungi and bacteria differ significantly, thus the three-mediator system may not be suitable for selectively detecting similar bacterial strains in a mixed microbial suspension.”
1 of 1

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Authors
Smida, H., Langlard, A., Ameline, D., Thobie-Gautier, C., Boujtita, M., & Lebègue, E.
Affiliations
Centre National de la Recherche Scientifique (FR)
Corresponding author
Lebègue, Estelle
ORCID
Venue
Analytical and Bioanalytical Chemistry
Publisher
Springer Science and Business Media LLC
Published
2023-02-09
Volume
415
Issue
18
Pages
3717-3725
ISSN
  • 1618-2642 (ISSN-L)
  • 1618-2650
DOI
10.1007/s00216-023-04568-z
OpenAlex
W4319657220
Cited by
22
References
44
Citations by year
2026: 4 · 2025: 5 · 2024: 10 · 2023: 3
FWCI
1.62
Type
journal article
Language
en
Open access
Licence
Locations
  • Analytical and Bioanalytical Chemistry (publishedVersion)
  • PubMed (publishedVersion)
  • HAL (Le Centre pour la Communication Scientifique Directe) (submittedVersion)
Funders
  • Conseil Régional des Pays de la Loire (e-NANOBIO)
  • Nantes Université
Topics
  • Physical Sciences › Engineering › Electrical and Electronic Engineering › Electrochemical sensors and biosensors
  • Physical Sciences › Chemistry › Electrochemistry › Electrochemical Analysis and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
Keywords
bacterial detection, chronoamperometry, ultramicroelectrodes, redox mediators, bacterial strain identification, virulence factors, pathogenic bacteria
MeSH terms
Bacteria, Electrochemistry, Oxidation-Reduction, Biosensing Techniques, Electrochemical Techniques

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