The recent development of nanoscale probes has enabled the study of single molecules and single cells with unprecedented resolution and the expansion of the field of single-entity electrochemistry. There is a growing evidence suggesting that highly crowded intracellular environment facilitate nanoelectrochemical measurements in cells by improving the signal-to-noise ratio. In this opinion piece, we discuss the concept of macromolecular crowding and its implications in single-entity electrochemistry. • The macromolecular crowding of the intracellular environment plays a crucial role in regulating cellular physiology. • Single-entity electrochemistry is enabling high-resolution intracellular measurements. • In vitro macromolecular crowding enhances the detection of single entities.