PASTEUR, Département de Chimie, Ecole Normale Supérieure, PSL University, Sorbonne Université, CNRS, 75005 Paris, France.
University of Bordeaux, CNRS, Bordeaux INP, CBMN, UMR 5248 CNRS, F-33600 Pessac, France.
Anal Chem. 2022 Jan 25;94(3):1686-1696. doi: 10.1021/acs.analchem.1c04238. Epub 2022 Jan 7.
In this work, the characterization of release events from liposomes has been addressed quantitatively by an electrochemiluminescence (ECL) imaging strategy. First, ECL reagents ([Ru(bpy)] and tripropylamine) were encapsulated in sealed giant asymmetrical liposomes (100 μm in diameter) made of DOPG/DOPC phospholipids. After sedimentation on an indium tin oxide electrode material, the opening of liposomes was triggered by polarization of the surface. Under these conditions, amperometry, epifluorescence imaging, and ECL imaging were combined and synchronized to monitor and image the rupture of giant liposomes during the release and subsequent ECL emission of their redox content. Amperometry allowed the quantification of the content released from single liposomes. The location and status of liposomes (closed or opened) were assessed by epifluorescence imaging. ECL provided the image of the efflux of matter after liposome opening. This original ECL imaging approach favorably compares with strictly photoluminescent or electrochemical techniques and appears to be adapted for the investigation of membrane rupture/permeation events.
在这项工作中,通过电化学发光(ECL)成像策略对脂质体的释放事件进行了定量表征。首先,将 ECL 试剂([Ru(bpy)]和三丙胺)封装在由 DOPG/DOPC 磷脂制成的密封的非对称巨型脂质体(直径 100μm)中。在沉积在氧化铟锡电极材料上之后,通过表面极化触发脂质体的开口。在这些条件下,将安培法、荧光成像和 ECL 成像相结合并同步,以监测和成像在释放过程中以及随后的氧化还原内容物的 ECL 发射期间巨型脂质体的破裂。安培法允许从单个脂质体中定量释放的内容物。通过荧光成像评估脂质体的位置和状态(闭合或打开)。ECL 提供了脂质体打开后的物质流出的图像。这种原始的 ECL 成像方法与严格的光致发光或电化学技术相比具有优势,并且似乎适用于研究膜破裂/渗透事件。