High Content Imaging Approaches for In Vitro Toxicology

Abstract

High Content Imaging (HCI) is a quantitative automated fluorescence microscopy approach which uses advanced algorithms for rapid analysis of images acquired from fluorescently stained cells. The term “High Content” derives from the fact that HCI allows for multiplexing of various readouts by use of several fluorescence probes at different wavelengths. This method facilitates information generation on multiple simultaneous readouts, for instance on morphological structures, spatial and dynamic processes of molecules, signal transduction, or enzyme activities. Moreover, mechanisms of compound-induced toxicity and the specific cellular pathways involved may be studied by combining the HCI approach with the use of specific enzyme inhibitors, enzyme inducers or RNA interference. As HCI provides multiplexed detailed information at the level of a single cell, as well as characterization of cellular population distributions, it offers a superior investigational tool compared to standard spectrophotometric plate readers that measure only average properties of a cell population.