In this article, ionic liquid-functionalized carbon nanodots (IL-CDs) were produced in a simple manner by electrochemical exfoliation of graphite rods in the presence of an amino-terminated ionic liquid, and their preliminary applications were exploited. TEM and AFM results showed that these IL-CDs are about 2.6 nm in diameter. The small-sized IL-CDs have strong photoluminescence, with a quantum yield of about 11.3%, and could be used for cell imaging. Moreover, the IL-CDs exhibit good electron transfer properties and catalytic activities for O-2 and H2O2 reduction. Additionally, the as-prepared IL-CDs can be applied as a matrix for immobilizing enzymes (glucose oxidase) to construct biosensors. Due to these favorable properties, IL-CDs will find promising practical applications in electrocatalysis, biosensing, and bioimaging.