University of Bordeaux , Bordeaux INP, ISM, UMR CNRS 5255 , 33607 Pessac , France.
Department of Chemistry "G. Ciamician" , University of Bologna , Via Selmi 2 , 40126 Bologna , Italy.
J Am Chem Soc. 2018 Nov 7;140(44):14753-14760. doi: 10.1021/jacs.8b08080. Epub 2018 Oct 29.
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.
本文报道了一种基于电致化学发光(ECL)的表面限制显微镜,该显微镜允许在与电极的界面处对单个细胞的质膜进行成像。通过分析哺乳动物 CHO 细胞的光致发光(PL)、ECL 和 AFM 图像,我们证明与宽场荧光不同,ECL 发射被限制在电极表面的紧邻区域,只有细胞的基底膜才会发光。所得到的 ECL 显微镜揭示了经典荧光显微镜无法分辨的细节,而无需任何光照射和特定设置。由于涉及短寿命电生成自由基的独特 ECL 机制,ECL 发射区域的厚度约为 500nm。此外,所报道的 ECL 显微镜是一种动态技术,它反映了通过细胞膜的传输特性,而不仅仅是对细胞膜的特定标记。最后,使用喷墨打印在普通显微镜玻璃盖玻片上的基于一次性透明碳纳米管(CNT)的电极在反射和透射两种配置下用于对细胞成像。因此,我们的方法为 ECL 作为一种表面限制显微镜开辟了新的途径,以开发单细胞分析并对细胞或膜中的生物实体的动态进行成像。