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Evaluation of a high‐content screening fluorescence‐based assay analyzing the pharmacological modulation of lipid homeostasis in human macrophages
Author(s) -
Werner Tobias,
Liebisch Gerhard,
Grandl Margot,
Schmitz Gerd
Publication year - 2006
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20237
Subject(s) - phospholipid , cholesterol , staining , fluorescence , fluorescent staining , efflux , ethanolamine , chemistry , flow cytometry , biochemistry , biology , microbiology and biotechnology , membrane , genetics , physics , quantum mechanics
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