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 develope...
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| Published in | Cytometry. Part A Vol. 69A; no. 3; pp. 200 - 202 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
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| Abstract | 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, 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 |
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| AbstractList | 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, 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 Abstract 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 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. 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. 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. BACKGROUNDFor 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.METHODSMaking 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.RESULTSMorphologic 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. |
| Author | Grandl, Margot Schmitz, Gerd Liebisch, Gerhard Werner, Tobias |
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| Cites_doi | 10.1034/j.1600-0854.2002.030404.x 10.1093/emboj/19.5.892 10.1161/01.ATV.18.9.1424 10.1016/S0022-2275(20)32205-7 |
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| Notes | Part of this work was presented at the 10th Leipziger Workshop “Systems Biology and Clinical Cytomics,” April 7–9, 2005, Leipzig, Germany. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
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| References | 1998; 18 2002; 3 2000; 19 2001; 42 e_1_2_5_3_2 e_1_2_5_2_2 e_1_2_5_5_2 e_1_2_5_4_2 |
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For understanding cholesterol and phospholipid efflux pathways there is a need for cellular fluorescence‐based high‐content screens (HCS) to... For understanding cholesterol and phospholipid efflux pathways there is a need for cellular fluorescence-based high-content screens (HCS) to investigated the... Abstract Background For understanding cholesterol and phospholipid efflux pathways there is a need for cellular fluorescence‐based high‐content screens (HCS)... BACKGROUNDFor understanding cholesterol and phospholipid efflux pathways there is a need for cellular fluorescence-based high-content screens (HCS) to... |
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| SubjectTerms | Cell Membrane - drug effects Cell Membrane - metabolism Cholesterol - pharmacology fluorescence Fluorescent Dyes - chemistry high‐content screening Homeostasis - drug effects Humans Image Cytometry - methods lipid homeostasis 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 |
| Title | Evaluation of a high‐content screening fluorescence‐based assay analyzing the pharmacological modulation of lipid homeostasis in human macrophages |
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