Stimuli‐Responsive DNA Nanomachines for Intracellular Targeted Electrochemiluminescence Imaging in Single Cells

Abstract Electrochemiluminescence (ECL) microscopy has emerged as a powerful technique for single‐cell imaging owing to its unparalleled background‐free imaging advantages. However, controlled intracellular ECL imaging remains challenging. Here, we developed a stimuli‐responsive self‐assembled DNA nanomachine that enables the ECL imaging of intracellular target biomolecules in single cells. The ECL nanoprobe consists of an ECL nanoemitter constructed from Ru(bpy) 3 2+ ‐doped metal–organic framework as the nanoreactor core, with a DNA polymer hydrogel (DNAgel) shell acting as the stimuli‐gated layer. The outer functionalized DNAgel of the ECL nanoprobe was specifically designed to block ECL generation and to dissociate by ATP molecules, thereby enabling selective recovery of ECL emission capability. Such an engineered stimuli‐responsive nanomachine successfully achieved the targeted ECL imaging of intracellular ATP distribution with spatial resolution. In addition, ECL imaging of various intracellular biomolecules should be generalizable by simply changing the switching DNA sequence of the probe. Our research provided a reliable strategy for ECL microscopy within cells, which will broaden the application of ECL in single‐cell and single‐molecule profiling.