Single-Entity Electrochemistry for Digital Biosensing at Ultralow Concentrations

[Image: see text] Quantifying ultralow analyte concentrations is a continuing challenge in the analytical sciences in general and in electrochemistry in particular. Typical hurdles for affinity sensors at low concentrations include achieving sufficiently efficient mass transport of the analyte, dealing with slow reaction kinetics, and detecting a small transducer signal against a background signal that itself fluctuates slowly in time. Recent decades have seen the advent of methods capable of detecting single analytes ranging from the nanoscale to individual molecules, representing the ultimate mass sensitivity to these analytes. However, single-entity detection does not automatically translate into a superior concentration sensitivity. This is largely because electrochemical transducers capable of such detection are themselves miniaturized, exacerbating mass transport and binding kinetic limitations. In this Perspective, we discuss how these challenges can be tackled through so-called digital sensing: large arrays of separately addressable single-entity detectors that provide real-time information on individual binding events. We discuss the advantages of this approach and the barriers to its implementation.