Phospholipidosis detection

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(New page: = Phospholipidosis detection in human macrophages using NBD-PE = <br>Four day M-CSF differentiated macrophages were cultured 24h with E-LDL or Ox-LDL and 12g/ml of the fluorescently t...)
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= Phospholipidosis detection in human macrophages using NBD-PE  =
= Phospholipidosis detection in human macrophages using NBD-PE  =
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<br>Four day M-CSF differentiated macrophages were cultured 24h with E-LDL or Ox-LDL and 12g/ml of the fluorescently tagged phospholipid, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (NBD-PE) (Molecular Probes, Eugene, OR, USA). Amiodarone (15μmol/L) and amitryptilin (30μmol/L) were used as a positive control. The final concentration of DMSO vehicle was 0.3% which does not affect NBD-PE accumulation of cytotoxicity results.&nbsp; Afterwards cells were fixed with 4% paraformaldehyde for 20 min at room temperature and washed three times with PBS. The method was adapted from Morelli JK et al Cell Biol Toxicol 22 (2006) 15-27.<br>
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<br>Four day M-CSF differentiated macrophages were cultured 24h with E-LDL or Ox-LDL and 12µg/ml of the fluorescently tagged phospholipid, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (NBD-PE) (Molecular Probes, Eugene, OR, USA). Amiodarone (15μmol/L) and amitryptilin (30μmol/L) were used as a positive control. The final concentration of DMSO vehicle was 0.3% which does not affect NBD-PE accumulation of cytotoxicity results.&nbsp; Afterwards cells were fixed with 4% paraformaldehyde for 20 min at room temperature and washed three times with PBS. The method was adapted from Morelli JK et al Cell Biol Toxicol 22 (2006) 15-27.<br>
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[[Category:Lipid imaging]]

Revision as of 15:37, 22 June 2009

Phospholipidosis detection in human macrophages using NBD-PE


Four day M-CSF differentiated macrophages were cultured 24h with E-LDL or Ox-LDL and 12µg/ml of the fluorescently tagged phospholipid, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (NBD-PE) (Molecular Probes, Eugene, OR, USA). Amiodarone (15μmol/L) and amitryptilin (30μmol/L) were used as a positive control. The final concentration of DMSO vehicle was 0.3% which does not affect NBD-PE accumulation of cytotoxicity results.  Afterwards cells were fixed with 4% paraformaldehyde for 20 min at room temperature and washed three times with PBS. The method was adapted from Morelli JK et al Cell Biol Toxicol 22 (2006) 15-27.

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