Evaluation of a high‐content screening fluorescence‐based assay analyzing the pharmacological modulation of lipid homeostasis in human macrophages

Background For understanding cholesterol and phospholipid efflux pathways there is a need for cellular fluorescence‐based high‐content screens (HCS) to investigated the cholesterol and phospholipid content in human macrophages. Methods Making use of fluorescence imaging based on HCS we have developed a tool to evaluate new agents that can act as inducers of cholesterol efflux. The fluorescence assay is based on the different staining patterns of cholesterol‐loaded (E‐LDL) and deloaded (HDL 3 ) differentiated monocytes by the saturated, fluorescene lipid probe (1,2‐dimyristoyl‐sn‐glycero‐3‐phospho‐ethanolamine)‐tetramethyl‐rhodamin. Results Morphologic examination and statistical evaluation of the staining pattern such as gray value, threshold area, shape factor and the spot size distribution provides evidence for a significant pattern change when cholesterol enriched and cholesterol depleted differentiated monocytes were imaged. © 2006 International Society for Analytical Cytology