Cell‐based biosensors (
CBB
s) utilizing impedance measurement have become a powerful tool for cytotoxicity analysis of (i) engineered micro‐ and nanoparticles (NPs) and (ii) complex mixtures of environmental particulate matter (
PM
). With the recent increase in the development and application of NPs, bio‐analytical techniques capable of fast, reliable, and accurate cytotoxicity analysis are needed to prioritize these materials for further toxicological testing to ensure their safe use, both for human health and environmental safety. This article focuses on the development and application of impedance‐based
CBBs
for rapid, sensitive, efficient, and label‐free analysis of micro‐ and nanoparticle‐induced cytotoxicity through the monitoring of several cellular responses, including cell adhesion, spreading, proliferation, transepithelial or trans endothelial electrical resistance (
TER
), and micromotion. In addition, these techniques are potentially useful for air quality monitoring, as demonstrated through the cytotoxicity analysis of complex mixtures of PM. Instruments based on electric cell–substrate impedance sensing (
ECIS
) and real‐time cell analysis (
RTCA
) are the two most widely used impedance‐based devices commercially available. Thus, detailed descriptions of the principles directing the impedance‐based measurements and data analysis employed in these systems are presented.