Cell-Based Imaging Assay for Detection of Phospholipidosis.

Accumulation of lysosomal phospholipids in cells exposed to cationic amphiphilic drugs is characteristic of drug-induced phospholipidosis. The morphological hallmark of phospholipidosis is the appearance of unicentric or multicentric-lamellar bodies when viewed under an electron microscope (EM). The EM method, the gold standard of detecting cellular phospholipidosis, has downsides, namely, low-throughput, high-costs, and unsuitability for screening a large chemical library. This chapter describes a cell-based high-content phospholipidosis assay using the LipidTOX reagent in a high-throughput screening (HTS) platform. This assay has been optimized and validated in HepG2 and HepRG cells, and miniaturized into a 1536-well plate, thus can be used for high-throughput screening (HTS) to identify chemical compounds that induce phospholipidosis.