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

Cytometry A. 2006 Mar;69(3):200-2. doi: 10.1002/cyto.a.20237.


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 (HDL3) differentiated monocytes by the saturated, fluorescent 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.

Publication types

  • Evaluation Study

MeSH terms

  • Cell Membrane / drug effects
  • Cell Membrane / metabolism
  • Cholesterol / pharmacology
  • Fluorescent Dyes / chemistry
  • Homeostasis / drug effects
  • Humans
  • Image Cytometry / methods*
  • Lipid Metabolism / drug effects*
  • Lipoproteins, HDL / pharmacology
  • Lipoproteins, HDL3
  • Lipoproteins, LDL / chemistry
  • Lipoproteins, LDL / pharmacology
  • Macrophages / cytology
  • Macrophages / drug effects
  • Macrophages / metabolism*
  • Membrane Microdomains / drug effects*
  • Membrane Microdomains / metabolism
  • Phosphatidylethanolamines / chemistry
  • Reproducibility of Results
  • Rhodamines / chemistry


  • Fluorescent Dyes
  • Lipoproteins, HDL
  • Lipoproteins, HDL3
  • Lipoproteins, LDL
  • Phosphatidylethanolamines
  • Rhodamines
  • tetramethylrhodamine
  • Cholesterol
  • 1,2-dimyristoylphosphatidylethanolamine