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We report here the development of coreactant-based electrogenerated chemiluminescence (ECL) as a surface-confined microscopy to image single cells and their membrane proteins. Labeling the entire cell membrane allows one to demonstrate that, by contrast with fluorescence, ECL emission is only detected from fluorophores located in the immediate vicinity of the electrode surface (i.e., 1–2 μm). Then, to present the potential diagnostic applications of our approach, we selected carbon nanotubes (CNT)-based inkjet-printed disposable electrodes for the direct ECL imaging of a labeled plasma receptor overexpressed on tumor cells. The ECL fluorophore was linked to an antibody and enabled to localize the ECL generation on the cancer cell membrane in close proximity to the electrode surface. Such a result is intrinsically associated with the unique ECL mechanism and is rationalized by considering the limited lifetimes of the electrogenerated coreactant radicals. The electrochemical stimulus used for luminescence generation does not suffer from background signals, such as the typical autofluorescence of biological samples. The presented surface-confined ECL microscopy should find promising applications in ultrasensitive single cell imaging assays.

Polyacrylamide-coated, carbon nanotube (PA/CNT) electrodes were prepared by an inkjet printing process and used to measure pyocyanin and uric acid in a wound fluid simulant at 37 °C. These two molecules are potential indicators of infection, and therefore their detection could prove useful for monitoring wound healing. Pyocyanin is a marker for the common wound bacterium Pseudomonas aeruginosa. Our long-term goal is to use these inexpensive and disposable electrodes to measure biomarkers of wound healing directly. In this proof-of-concept work, studies were performed in a wound fluid simulant to evaluate the stability of the electrodes and their responsiveness for the two bioanalytes. The PA/CNT inkjet-printed electrodes and electrical contacts were stable with unchanging physical and electrochemical properties in the wound fluid simulant over a 7–8-day period at 37 °C. The detection figures of merit for pyocyanin in the simulant at 37 °C were as follows: linear over the physiologically relevant range = 0.10 to 100 μmol L–1(R2= 0.9992), limit of detection = 0.10 μmol L–1(S/N = 3), sensitivity = 35.6 ± 0.8 mA-L mol–1and response variability ≤4% RSD. The detection figures of merit for uric acid in the simulant at 37 °C were as follows: linear over the physiologically relevant range = 100 to 1000 μmol L–1(R2= 0.9997), sensitivity = 2.83 ± 0.01 mA-L mol–1, and response variability ≤4% RSD. The limit of detection was not determined. The PA/CNT electrodes were also used to quantify pyocyanin concentrations in cell-free culture media from different strains of P. aeruginosa. The detected concentrations ranged from 1.00 ± 0.02 to 118 ± 6 μM depending on the strain.

Herein is reported a surface-confined microscopy based on electrochemiluminescence (ECL) that allows to image the plasma membrane of single cells at the interface with an electrode. By analyzing photoluminescence (PL), ECL and AFM images of mammalian CHO cells, we demonstrate that, in contrast to the wide-field fluorescence, ECL emission is confined to the immediate vicinity of the electrode surface and only the basal membrane of the cell becomes luminescent. The resulting ECL microscopy reveals details that are not resolved by classic fluorescence microscopy, without any light irradiation and specific setup. The thickness of the ECL-emitting regions is ∼500 nm due to the unique ECL mechanism that involves short-lifetime electrogenerated radicals. In addition, the reported ECL microscopy is a dynamic technique that reflects the transport properties through the cell membranes and not only the specific labeling of the membranes. Finally, disposable transparent carbon nanotube (CNT)-based electrodes inkjet-printed on classic microscope glass coverslips were used to image cells in both reflection and transmission configurations. Therefore, our approach opens new avenues for ECL as a surface-confined microscopy to develop single cell assays and to image the dynamics of biological entities in cells or in membranes.

This work describes a new sensitive sensor for the simultaneous electrochemical determination by square wave anodic stripping voltammetry of Cd2+and Pb2+ions in aqueous solution, based on a disposable ink-jet-printed graphene electrode (IPGE) modified by a thin film of montmorillonite (Mont) clay mineral. The clay was characterized by X-ray diffraction and scanning electron microscopy while the Mont–IPGE was analyzed by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry before its exploitation for sensing studies. EIS results revealed a low electron transfer rate on Mont–IPGE during the electro-oxidation of [Fe(CN)6]3−. The stripping response on both Cd2+and Pb2+analytes was improved by the in situ co-deposition with bismuth, added to the electrolyte solution, on the clay substrate. The proposed electrode showed great stability and good reproducibility. The key experimental parameters governing the stripping analysis of Cd2+and Pb2+were optimized. A linear relationship between peak currents and concentrations of the analytes was obtained in the range from 0.01 to 0.21 µM, leading to detection limits of 0.42 nM for Cd2+and 1.14 nM for Pb2+ions, respectively (S/N= 3). The sensitivities to Cd2+and Pb2+ions were 921.8 A cm−2M−1and 1960.8 A cm−2M−1, respectively. The Mont–IPGE sensor was used for the quantification of lead and cadmium in a mineral water.

Multi-layer inkjet printing is applied to prepare microchips that consist of up to eight three-electrode sensors comprising a carbon nanotube (CNT) working electrode, a silver/silver chloride quasi-reference electrode and a CNT counter electrode. Optionally, the sensors contain micro-wells to measure sample volumes as small as 50 µL. The implementation of inkjet printing allows the flexible design and highly reproducible fabrication of batches of various electrode shapes and dimensions. The usage of CNTs as stand-alone working electrodes results in electroanalytical sensors with reliable and superior electrochemical response compared to tested commercial carbon paste electrodes. The sensors can be used at least up to 20 times, e.g., based on an immunoassay based strategy, or in a disposable way, e.g., when measuring in complex matrices such as blood. A gold ink is applied as alternative working electrode material. The printed sensors can be used with compact multichannel potentiostats that are suitable for point-of-care diagnostics.