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.
In the present work we numerically simulated the electrogenerated chemiluminescence (ECL) from a Ru(bpy)3 2+-doped silica nanoparticle (Ru-DSNP) in buffer containing tripropylamine (TPrA). An experimental study reported from Zanarini et al. showed that ECL intensity for the Ru-DSNP/TPrA system exhibits two emission waves, while the potential of the working electrode is swept in the positive direction. The first ECL wave with a peak at ∼0.9 V (vs Ag|AgCl) is triggered by TPrA oxidation and is governed by the deprotonation equilibrium of TPrA cation radical (TPrA•+ = TPrA• + H+). We present a model for the description of the first ECL wave, which also takes into consideration the influence on the deprotonation equilibrium of the electrode surface functionality. This model indicated that the detachment of a Ru-DSNP (initially bound to the electrode surface via alkylthiols linkers) from the electrode surface and the subsequent electrode surface oxidation facilitate the radical deprotonation on the electrode surface causing the ECL quenching. The second ECL wave having its peak at ∼1.2 V is triggered by direct Ru(bpy)3 2+ oxidation. We modeled the second ECL wave as related to the electron hopping mechanism between Ru(bpy)3 2+ labels inside the Ru-DSNP. The results of the numerical simulations indicate that electrode surface functionality modification, which occurs during potential sweep, and the electron hopping mechanism between Ru(bpy)3 2+ labels play important roles in defining the Ru-DSNP/TPrA ECL signal.
Bipolar electrochemistry has been regarded as a powerful and sustainable electrochemical process for the synthesis of novel functional materials. The appealing features of this electrochemical technology, such as the wireless nature of the bipolar electrode (BPE) and the possibility to drive simultaneously electrochemical reactions on multiple BPEs placed in the same electrochemical cell, together with the possibility to change the shape and positioning of the driving electrodes, give significant freedom to design reaction systems. Nevertheless, the cell geometry dramatically affects the distribution and intensity of the potential gradient generated on the BPE surface and its monitoring is hampered due to the wireless nature of the BPE. In the present study, we propose the use of electrochemiluminescence (ECL) as an electrochemical imaging technique to map the distribution of potential gradient in bipolar electrochemical cells with different geometries. The proposed approach exploits the strong ECL emission of luminol/hydrogen peroxide (H2O2) system generated at the anodic pole of the BPE, when the total applied voltage (E tot) is strong enough to trigger the electrochemical reaction. Since luminol ECL emission is rather intense and relatively stable, the evolution of the potential distribution as a function of E tot can be monitored using a digital camera, allowing the elucidation of the potential distribution profile in every bipolar configuration. The suggested approach represents a valuable and reliable method to map the potential gradient in bipolar electrochemical systems and can be readily employed in every type of bipolar configuration.
Mammalian cell analysis is essential in the context of both fundamental studies and clinical applications. Among the various techniques available for cell analysis, electrochemiluminescence (ECL) has attracted significant attention due to its integration of both electrochemical and spectroscopic methods. In this review, we summarize recent advances in the ECL-based systems developed for mammalian cell analysis. The review begins with a summary of the developments in luminophores that opened the door to ECL applications for biological samples. Secondly, ECL-based imaging systems are introduced as an emerging technique to visualize single-cell morphologies and intracellular molecules. In the subsequent section, the ECL sensors developed in the past decade are summarized, the use of which made the highly sensitive detection of cell-derived molecules possible. Although ECL immunoassays are well developed in terms of commercial use, the sensing of biomolecules at a single-cell level remains a challenge. Emphasis is therefore placed on ECL sensors that directly detect cellular molecules from small portions of cells or even single cells. Finally, the development of bipolar electrode devices for ECL cell assays is introduced. To conclude, the direction of research in this field and its application prospects are described.
Monitoring Prostate Cancer (PCa) biomarkers is an efficient way to diagnosis this disease early, since it improves the therapeutic success rate and suppresses PCa patient mortality: for this reason a powerful analytical technique such as electrochemiluminescence (ECL) is already used for this application, but its widespread usability is still hampered by the high cost of commercial ECL equipment. We describe an innovative approach for the selective and sensitive detection of the PCa biomarker sarcosine, obtained by a synergistic ECL-supramolecular approach, in which the free base form of sarcosine acts as co-reagent in a Ru(bpy)3(2+)-ECL process. We used magnetic micro-beads decorated with a supramolecular tetraphosphonate cavitand (Tiiii) for the selective capture of sarcosine hydrochloride in a complex matrix like urine. Sarcosine determination was then obtained with ECL measurements thanks to the complexation properties of Tiiii, with a protocol involving simple pH changes - to drive the capture-release process of sarcosine from the receptor - and magnetic micro-bead technology. With this approach we were able to measure sarcosine in the μM to mM window, a concentration range that encompasses the diagnostic urinary value of sarcosine in healthy subjects and PCa patients, respectively. These results indicate how this ECL-supramolecular approach is extremely promising for the detection of sarcosine and for PCa diagnosis and monitoring, and for the development of portable and more affordable devices.
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