RESUMO
The respiratory activity of cultured cells can be electrochemically monitored using scanning electrochemical microscopy (SECM) with high spatial resolution. However, in SECM, the electrode takes a long time to scan, limiting simultaneous measurements with large biological samples such as cell spheroids. Therefore, for rapid electrochemical imaging, a novel strategy is needed. Herein, we report electrochemiluminescence (ECL) imaging of spheroid respiratory activity for the first time using sequential potential steps. L-012, a luminol analog, was used as an ECL luminophore, and H2O2, a sensitizer for ECL of L-012, was generated by the electrochemical reduction of dissolved O2. The ECL imaging visualized spheroid respiratory activity-evidenced by ECL suppression-corresponding to O2 distribution around the spheroids. This method enabled the time-lapse imaging of respiratory activity in multiple spheroids with good spatial resolution comparable to that of SECM. Our work provides a promising high-throughput imaging strategy for elucidating spheroid cellular dynamics.
Assuntos
Técnicas Biossensoriais , Esferoides Celulares , Técnicas Eletroquímicas , Eletrodos , Peróxido de Hidrogênio , Medições Luminescentes , LuminolRESUMO
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.