Scanning electrochemical microscopy (SECM) is a powerful technique that can provide chemical identity, quantification, and spatiotemporal information on biosurfaces. The ability of SECM for noninvasive and high-resolution electrochemical imaging has made it valuable for the study of cell phenotypes and functions. This review focuses on the latest advances of SECM technique for the biosurface imaging. The SECM measurements of different biomarkers, including oxygen consumption rate and enzyme activity of cell aggregates, redox state of cardiomyocytes, and bacterial metabolic activity, are introduced. The applicability of SECM on membrane permeability measurements, neurotransmitter measurements, and intracellular measurements is discussed.