Shadow electrochemiluminescence imaging of giant liposomes opening at polarized electrodes.

In this work, the release of giant liposome (∼100 µm in diameter) content was imaged by shadow electrochemiluminescence (ECL) microscopy. Giant unilamellar liposomes were pre-loaded with a sucrose solution and allowed to sediment at an ITO electrode surface immersed in a solution containing a luminophore ([Ru(bpy)3]2+) and a sacrificial co-reactant (tri-n-propylamine). Upon polarization, the electrode exhibited illumination over its entire surface thanks to the oxidation of ECL reagents. However, as soon as liposomes reached the electrode surface, dark spots appeared and then spread over time on the surface. This observation reflected a blockage of the electrode surface at the contact point between the liposome and the electrode surface, followed by the dilution of ECL reagents after the rupture of the liposome membrane and release of its internal ECL-inactive solution. Interestingly, ECL reappeared in areas where it initially faded, indicating back-diffusion of ECL reagents towards the previously diluted area and thus confirming liposome permeabilization. The whole process was analyzed qualitatively and quantitatively within the defined region of interest. Two mass transport regimes were identified: a gravity-driven spreading process when the liposome releases its content leading to ECL vanishing and a diffusive regime when ECL recovers. The reported shadow ECL microscopy should find promising applications for the imaging of transient events such as molecular species released by artificial or biological vesicles.

Electrochemiluminescence Microscopy.

Electrochemiluminescence (ECL) is rapidly evolving from an analytical method into an optical microscopy. The orthogonality of the electrochemical trigger and the optical readout distinguishes it from classic microscopy and electrochemical techniques, owing to its near-zero background, remarkable sensitivity, and absence of photobleaching and phototoxicity. In this review, we summarize the recent advances in ECL imaging technology, emphasising original configurations which enable the imaging of biological entities and the improvement of the analytical properties by increasing the complexity and multiplexing of bioassays. Additionally, mapping the (electro)chemical reactivity in space provides valuable information on nanomaterials and facilitates deciphering ECL mechanisms for improving their performances in diagnostics and (electro)catalysis. Finally, we highlight the recent achievements in imaging at the ultimate limits of single molecules, single photons or single chemical reactions, and the current challenges to translate the ECL imaging advances to other fields such as material science, catalysis and biology.

Wireless rotating bipolar electrochemiluminescence for enzymatic detection.

New dynamic, wireless and cost-effective analytical devices are developing rapidly in biochemical analysis. Here, we report on a remotely-controlled rotating electrochemiluminescence (ECL) sensing system for enzymatic detection of a model analyte, glucose, on both polarized sides of an iron wire acting as a bipolar electrode. The iron wire is controlled by double contactless mode, involving remote electric field polarization, and magnetic field-induced rotational motion. The former triggers the interfacial polarization of both extremities of the wire by bipolar electrochemistry, which generates ECL emission of the luminol derivative (L-012) with the enzymatically produced hydrogen peroxide in presence of glucose, at both anodic and cathodic poles, simultaneously. The latter generates a convective flow, leading to an increase in mass transfer and amplifying the corresponding ECL signals. Quantitative glucose detection in human serum samples is achieved. The ECL signals were found to be a linear function of the glucose concentration within the range of 10-1000 µM and with a limit of detection of 10 µM. The dynamic bipolar ECL system simultaneously generates light emissions at both anodic and cathodic poles for glucose detection, which can be further applied to biosensing and imaging in autonomous devices.

Wireless Multimodal Light-Emitting Arrays Operating on the Principles of LEDs and ECL.

Electrochemistry-based light-emitting devices have gained considerable attention in different applications such as sensing and optical imaging. In particular, such systems are an interesting alternative for the development of multimodal light-emitting platforms. Herein we designed a multicolor light-emitting array, based on the electrochemical switch-on of light-emitting diodes (LEDs) with a different intrinsic threshold voltage. Thermodynamically and kinetically favored coupled redox reactions, i. e. the oxidation of Mg and the reduction of protons on Pt, act as driving force to power the diodes. Moreover, this system enables to trigger an additional light emission based on the interfacial reductive-oxidation electrochemiluminescence (ECL) mechanism of the Ru(bpy)3 2+/S2O8 2- system. The synergy between these light-emission pathways offers a multimodal platform for the straightforward optical readout of physico-chemical information based on composition changes of the solution.

Wireless Light-Emitting Electrode Arrays for the Evaluation of Electrocatalytic Activity.

Water splitting has become a sustainable and clean alternative for hydrogen production. Commonly, the efficiency of such reactions is intimately related to the physico-chemical properties of the catalysts that constitute the electrolyzer. Thus, the development of simple and fast methods to evaluate the electrocatalytic efficiency of an electrolyzer is highly required. In this work, we present an unconventional method based on the combination of bipolar electrochemistry and light-emitting diodes, which allows the evaluation of the electrocatalytic performance of the two types of catalysts, composing an electrolyzer, namely for oxygen and hydrogen evolution reactions, respectively. The integrated light emission of the diode acts as an optical readout of the electrocatalytic information, which simultaneously depends on the composition of the anode and the cathode. The electrocatalytic activity of Au, Pt, and Ni electrodes, connected to the LED in multiple anode/cathode configurations, towards the water splitting reactions has been evaluated. The efficiency of the electrolyzer can be represented in terms of the onset electric field (ϵonset) for light emission, obtaining variations that are in agreement with data reported with conventional electrochemistry. This work introduces a straightforward method for evaluating electrocatalysts and underscores the importance of material characterization in developing efficient electrolyzers for hydrogen production.

Infrared photoinduced electrochemiluminescence microscopy of single cells.

Electrochemiluminescence (ECL) is evolving rapidly from a purely analytical technique into a powerful microscopy. Herein, we report the imaging of single cells by photoinduced ECL (PECL; λem = 620 nm) stimulated by an incident near-infrared light (λexc = 1050 nm). The cells were grown on a metal-insulator-semiconductor (MIS) n-Si/SiOx/Ir photoanode that exhibited stable and bright PECL emission. The large anti-Stokes shift allowed for the recording of well-resolved images of cells with high sensitivity. PECL microscopy is demonstrated at a remarkably low onset potential of 0.8 V; this contrasts with classic ECL, which is blind at this potential. Two imaging modes are reported: (i) photoinduced positive ECL (PECL+), showing the cell membranes labeled with the [Ru(bpy)3]2+ complex; and (ii) photoinduced shadow label-free ECL (PECL-) of cell morphology, with the luminophore in the solution. Finally, by adding a new dimension with the near-infrared light stimulus, PECL microscopy should find promising applications to image and study single photoactive nanoparticles and biological entities.

Near-IR Photoinduced Electrochemiluminescence Imaging with Structured Silicon Photoanodes.

Infrared (IR) imaging devices that convert IR irradiation (invisible to the human eye) to a visible signal are based on solid-state components. Here, we introduce an alternative concept based on light-addressable electrochemistry (i.e., electrochemistry spatially confined under the action of a light stimulus) that involves the use of a liquid electrolyte. In this method, the projection of a near-IR image (λexc = 850 or 840 nm) onto a photoactive Si-based photoanode, immersed into a liquid phase, triggers locally the photoinduced electrochemiluminescence (PECL) of the efficient [Ru(bpy)3]2+-TPrA system. This leads to the local conversion of near-IR light to visible (λPECL = 632 nm) light. We demonstrate that compared to planar Si photoanodes, the use of a micropillar Si array leads to a large enhancement of local light generation and considerably improves the resolution of the PECL image by preventing photogenerated minority carriers from diffusing laterally. These results are important for the design of original light conversion devices and can lead to important applications in photothermal imaging and analytical chemistry.

Electrochemiluminescent imaging of a NADH-based enzymatic reaction confined within giant liposomes.

Herein, transient releases either from NADH-loaded liposomes or enzymatic reactions confined in giant liposomes were imaged by electrochemiluminescence (ECL). NADH was first encapsulated with the [Ru(bpy)3]2+ luminophore inside giant liposomes (around 100 µm in diameter) made of DOPC/DOPG phospholipids (i.e., 1,2-dioleolyl-sn-glycero-3-phosphocholine/1,2-dioleoyl-sn-glycerol-3-phospho-(1'-rac-glycerol) sodium salt) on their inner- and outer-leaflet, respectively. Then, membrane permeabilization triggered upon contact between the liposome and a polarized ITO electrode surface and ECL was locally generated. Combination of amperometry, photoluminescence, and ECL provided a comprehensive monitoring of a single liposome opening and content release. In a second part, the work is focused on the ECL characterization of NADH produced by glucose dehydrogenase (GDH)-catalyzed oxidation of glucose in the confined environment delimited by the liposome membrane. This was achieved by encapsulating both the ECL and catalytic reagents (i.e., the GDH, glucose, NAD+, and [Ru(bpy)3]2+) in the liposome. In accordance with the results obtained, NADH can be used as a biologically compatible ECL co-reactant to image membrane permeabilization events of giant liposomes. Under these conditions, the ECL signal duration was rather long (around 10 s). Since many enzymatic reactions involve the NADH/NAD+ redox couple, this work opens up interesting prospects for the characterization of enzymatic reactions taking place notably in artificial cells and in confined environments.

Redox-mediated electrochemiluminescence enhancement for bead-based immunoassay.

Electrochemiluminescence (ECL) is a highly sensitive mode of detection utilised in commercialised bead-based immunoassays. Recently, the introduction of a freely diffusing water-soluble Ir(iii) complex was demonstrated to enhance the ECL emission of [Ru(bpy)3]2+ labels anchored to microbeads, but a comprehensive investigation of the proposed 'redox-mediated' mechanism was not carried out. In this work, we select three different water-soluble Ir(iii) complexes by virtue of their photophysical and electrochemical properties in comparison with those of the [Ru(bpy)3]2+ luminophore and the TPrA co-reactant. A systematic investigation of the influence of each Ir(iii) complex on the emission of the Ru(ii) labels on single beads by ECL microscopy revealed that the heterogeneous ECL can be finely tuned and either enhanced up to 107% or lowered by 75%. The variation of the [Ru(bpy)3]2+ ECL emission was correlated to the properties of each Ir(iii)-based mediator, which enabled us to decipher the mechanism of interaction and define guidelines for the future design of novel Ir(iii) complexes to further enhance the ECL emission of bead-based immunoassays. Ultimately, we showcase the potential of this technology for practical sample analysis in commercial instruments by assessing the enhancement of the collective ECL intensity from a bead-based system.

Modulation of circularly polarized luminescence by swelling of microgels functionalized with enantiopure [Ru(bpy)3]2+ luminophores.

Chemoresponsive microgels functionalized with enantiomeric Δ- or Λ-[Ru(bpy)3]2+ showed tunable chiroptical properties upon swelling and shrinking. The tuning is triggered by a modulation of the local mobility of [Ru(bpy)3]2+ upon addition of fructose, controlling interactions and distances between [Ru(bpy)3]2+ and phenylboronic acid.

Off-Grid Electrogenerated Chemiluminescence with Customized p-i-n Photodiodes.

Electrochemiluminescence (ECL) is the generation of light induced by an electrochemical reaction, driven by electricity. Here, an all-optical ECL (AO-ECL) system is developped, which triggers ECL by the illumination of electrically autonomous "integrated" photoelectrochemical devices immersed in the electrolyte. Because these systems are made using small and cheap devices, they can be easily prepared and readily used by any laboratories. They are based on commercially available p-i-n Si photodiodes (≈1 € unit-1), coupled with well-established ECL-active and catalytic materials, directly coated onto the component leads by simple and fast wet processes. Here, a Pt coating (known for its high activity for reduction reactions) and carbon paint (known for its optimal ECL emission properties) are deposited at cathode and anode leads, respectively. In addition to its optimized light absorption properties, using the commercial p-i-n Si photodiode eliminates the need for a complicated manufacturing process. It is shown that the device can emit AO-ECL by illumination with polychromatic (simulated sunlight) or monochromatic (near IR) light sources to produce visible photons (425 nm) that can be easily observed by the naked eye or recorded with a smartphone camera. These low-cost off-grid AO-ECL devices open broad opportunities for remote photodetection and portable bioanalytical tools.

All-Optical Electrochemiluminescence at Metal-Insulator-Semiconductor Diodes.

Pt/InGa/n-Si/SiOx/Pt devices were prepared by using standard chemical and sputtering processes. These systems are diodes comprising a frontside photoactive metal-insulator-semiconductor (MIS) n-Si/SiOx/Pt junction and a backside Pt/InGa/n-Si Ohmic contact. Pt/InGa/n-Si/SiOx/Pt was first characterized by dark-solid-state electrical and impedance measurements. Then, each side of the device was investigated by electrochemical means in the dark and under near-IR illumination at 850 nm in the luminol-H2O2 electrochemiluminescence (ECL) electrolyte. The results suggested the possibility of triggering an all-optical ECL (AO-ECL) at Pt/InGa/n-Si/SiOx/Pt. This was confirmed by studying AO-ECL at the monolithic, all-integrated Pt/InGa/n-Si/SiOx/Pt device, immersed in the ECL electrolyte. The conversion process can occur with good stability and the intensity of the visible emission (440 nm) depends on tunable parameters such as the illumination power density, O2 concentration, or the concentration of added H2O2. These results are important for the next developments of AO-ECL in sensing and microscopy.

Wireless Magnetoelectrochemical Induction of Rotational Motion.

Electromagnetically induced rotation is a key process of many technological systems that are used in daily life, especially for energy conversion. In this context, the Lorentz force-induced deviation of charges is a crucial physical phenomenon to generate rotation. Herein, they combine the latter with the concept of bipolar electrochemistry to design a wireless magnetoelectrochemical rotor. Such a device can be considered as a wet analog of a conventional electric motor. The main driving force that propels this actuator is the result of the synergy between the charge-compensating ion flux along a bipolar electrode and an external magnetic field applied orthogonally to the surface of the object. The trajectory of the wirelessly polarized rotor can be controlled by the orientation of the magnetic field relative to the direction of the global electric field, producing a predictable clockwise or anticlockwise motion. Fine-tuning of the applied electric field allows for addressing conducting objects having variable characteristic lengths.

Luminescent Metal-Organic Frameworks for Electrochemiluminescent Detection of Water Pollutants.

The modern lifestyle has increased our utilization of pollutants such as heavy metals, aromatic compounds, and contaminants that are of rising concern, involving pharmaceutical and personal products and other materials that may have an important environmental impact. In particular, the ultimate results of the intense use of highly stable materials, such as heavy metals and chemical restudies, are that they turn into waste materials, which, when discharged, accumulate in environmental water bodies. In this context, the present review presents the application of metal-organic frameworks (MOFs) in electrochemiluminescent (ECL) sensing for water pollutant detection. MOF composites applied as innovative luminophore or luminophore carriers, materials for electrode modification, and the enhancement of co-reaction in ECL sensors have enabled the sensitive monitoring of some of the most common contaminants of emerging concern such as heavy metals, volatile organic compounds, pharmaceuticals, industrial chemicals, and cyanotoxins. Moreover, we provide future trends and prospects associated with ECL MOF composites for environmental sensing.

Optics Determines the Electrochemiluminescence Signal of Bead-Based Immunoassays.

Electrochemiluminescence (ECL) is an optical readout technique that is successfully applied for the detection of biomarkers in body fluids using microbead-based immunoassays. This technology is of utmost importance for in vitro diagnostics and thus a very active research area but is mainly focused on the quest for new dyes and coreactants, whereas the investigation of the ECL optics is extremely scarce. Herein, we report the 3D imaging of the ECL signals recorded at single microbeads decorated with the ECL labels in the sandwich immunoassay format. We show that the optical effects due to the light propagation through the bead determine mainly the spatial distribution of the recorded ECL signals. Indeed, the optical simulations based on the discrete dipole approximation compute rigorously the electromagnetic scattering of the ECL emission by the microbead and allow for reconstructing the spatial map of ECL emission. Thus, it provides a global description of the ECL chemical reactivity and the associated optics. The outcomes of this 3D imaging approach complemented by the optical modeling provide insight into the ECL optics and the unique ECL chemical mechanism operating on bead-based immunoassays. Therefore, it opens new directions for mechanistic investigations, ultrasensitive ECL bioassays, and imaging.

Dynamic Mapping of Electrochemiluminescence Reactivity in Space: Application to Bead-Based Assays.

As an electrochemical technique offering an optical readout, electrochemiluminescence (ECL) evolved recently into a powerful microscopy technique with the visualization of a wide range of microscopic entities. However, the dynamic imaging of transient ECL events did not receive intensive attention due to the limited number of electrogenerated photons. Here, the reaction kinetics of the model ECL bioassay system was revealed by dynamic imaging of single [Ru(bpy)3]2+-functionalized beads in the presence of the efficient tripropylamine coreactant. The time profile behavior of ECL emission, the variations of the ECL layer thickness, and the position of maximum ECL intensity over time were investigated, which were not achieved by static imaging in previous studies. Moreover, the dynamics of the ECL emission were confronted with the simulation. The reported dynamic ECL imaging allows the investigation of the ECL kinetics and mechanisms operating in bioassays and cell microscopy.

Endogenous and exogenous wireless multimodal light-emitting chemical devices.

Multimodal imaging is a powerful and versatile approach that integrates and correlates multiple optical modalities within a single device. This concept has gained considerable attention due to its potential applications ranging from sensing to medicine. Herein, we develop several wireless multimodal light-emitting chemical systems by coupling two light sources based on different physical principles: electrochemiluminescence (ECL) occurring at the electrode interface and a light-emitting diode (LED) switched on by an electrochemically triggered electron flow. Endogenous (thermodynamically spontaneous redox process) and exogenous (requiring an external power source) bipolar electrochemistry acts as a driving force to trigger both light emissions at different wavelengths. The results presented here interconnect optical imaging and electrochemical reactions, providing a novel and so far unexplored alternative to design autonomous hybrid systems with multimodal and multicolor optical readouts for complex bio-chemical systems.

Local reactivity of metal-insulator-semiconductor photoanodes imaged by photoinduced electrochemiluminescence microscopy.

Localized photoinduced electrochemiluminescence (PECL) is studied on photoanodes composed of Ir microbands deposited on n-Si/SiOx. We demonstrate that PECL microscopy precisely imaged the hole-driven heterogeneous photoelectrochemical reactivity. The method is promising for elucidating the local activity of photoelectrodes that are employed in solar energy conversion.

Tris(2,2'-bipyridyl)ruthenium (II) complex as a universal reagent for the fabrication of heterogeneous electrochemiluminescence platforms and its recent analytical applications.

In recent years, electrochemiluminescence (ECL) has received enormous attention and has emerged as one of the most successful tools in the field of analytical science. Compared with homogeneous ECL, the heterogeneous (or solid-state) ECL has enhanced the rate of the electron transfer kinetics and offers rapid response time, which is highly beneficial in point-of-care and clinical applications. In ECL, the luminophore is the key element, which dictates the overall performance of the ECL-based sensors in various analytical applications. Tris(2,2'-bipyridyl)ruthenium (II) complex, Ru(bpy)32+, is a coordination compound, which is the gold-standard luminophore in ECL. It has played a key role in translating ECL from a "laboratory curiosity" to a commercial analytical instrument for diagnosis. The aim of the present review is to provide the principles of ECL and classical reaction mechanisms-particularly involving the heterogeneous Ru(bpy)32+/co-reactant ECL systems, as well as the fabrication methods and its importance over solution-phase Ru(bpy)32+ ECL. Then, we discussed the emerging technology in solid-state Ru(bpy)32+ ECL-sensing platforms and their recent potential analytical applications such as in immunoassay sensors, DNA sensors, aptasensors, bio-imaging, latent fingerprint detection, point-of-care testing, and detection of non-biomolecules. Finally, we also briefly cover the recent advances in solid-state Ru(bpy)32+ ECL coupled with the hyphenated techniques.

All-Optical Electrochemiluminescence.

Electrochemiluminescence (ECL) is widely employed for medical diagnosis and imaging. Despite its remarkable analytical performances, the technique remains intrinsically limited by the essential need for an external power supply and electrical wires for electrode connections. Here, we report an electrically autonomous solution leading to a paradigm change by designing a fully integrated all-optical wireless monolithic photoelectrochemical device based on a nanostructured Si photovoltaic junction modified with catalytic coatings. Under illumination with light ranging from visible to near-infrared, photogenerated holes induce the oxidation of the ECL reagents and thus the emission of visible ECL photons. The blue ECL emission is easily viewed with naked eyes and recorded with a smartphone. A new light emission scheme is thus introduced where the ECL emission energy (2.82 eV) is higher than the excitation energy (1.18 eV) via an intermediate electrochemical process. In addition, the mapping of the photoelectrochemical activity by optical microscopy reveals the minority carrier interfacial transfer mechanism at the nanoscale. This breakthrough provides an all-optical strategy for generalizing ECL without the need for electrochemical setups, electrodes, wiring constraints, and specific electrochemical knowledge. This simplest ECL configuration reported so far opens new opportunities to develop imaging and wireless bioanalytical systems such as portable point-of-care sensing devices.