Phospholipidosis detection
From LipidomicsWiki
(Difference between revisions)
(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 12g/ml of the fluorescently t...) |
|||
Line 1: | Line 1: | ||
= Phospholipidosis detection in human macrophages using NBD-PE = | = 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 | + | <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. 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> |
+ | |||
+ | [[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.