title: Analysis of the fluorescence of monodansylcadaverine-positive cytoplasmic structures during 7-ketocholesterol-induced cell death.
abstract: OBJECTIVE
  To analyze multilamellar cytoplasmic structures by confocal laser scanning microscopy (CLSM) combined with factor analysis of biomedical image sequences (FAMIS).
  
  
  METHODS
  After treatment of U937 cells with 7-ketocholesterol (7-keto), cytoplasmic alterations were assessed with monodansylcadaverine (MDC). By ultraviolet excitation of a confocal laser scanning microscope (UV-CLSM), spectral sequences were performed to characterize 7-keto and MDC distribution inside cells. FAMIS was used to transform the image sequences in factor curves and images.
  
  
  RESULTS
  By UV-CLSM, 7-keto fluorescence was detected together with MDC, which revealed morphologic cytoplasmic changes in cells. The factor images obtained from confocal image sequences emphasized the view of these results. These data are in agreement with biochemical characterizations of MDC-positive structures.
  
  
  CONCLUSIONS
  The combined use of confocal microscopy and FAMIS allowed us to detect MDC-positive cytoplasmic structures in 7-keto-treated cells and to colocalize MDC and 7-keto distribution. This new method confirms the usefulness of MDC as a marker of oxysterol-induced cell death.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U494, Centre Hospitalier Universitaire Pitié-Salpêtrière, 75634 Paris, France. kahn@imed.jussieu.fr
authors.forename: Edmond
authors.initials: E
authors.lastname: Kahn
authors.forename: Anne
authors.initials: A
authors.lastname: Vejux
authors.forename: Gérard
authors.initials: G
authors.lastname: Lizard
authors.forename: Ginette
authors.initials: G
authors.lastname: Bessède
authors.forename: Frédérique
authors.initials: F
authors.lastname: Frouin
authors.forename: Céline
authors.initials: C
authors.lastname: Prunet
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Bernengo
authors.forename: Frédéric
authors.initials: F
authors.lastname: Brau
authors.forename: Andrew
authors.initials: A
authors.lastname: Todd-Pokropek
authors.forename: Philippe
authors.initials: P
authors.lastname: Gambert
mesh_terms: D001562:Benzimidazoles / Q000737:chemistry; D002103:Cadaverine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000494:pharmacology; D002232:Carbocyanines / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002467:Cell Nucleus / Q000737:chemistry / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D022083:Cytoplasmic Structures / Q000737:chemistry* / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007653:Ketocholesterols / Q000032:analysis / Q000494:pharmacology*; D008564:Membrane Potentials / Q000187:drug effects; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000737:chemistry / Q000187:drug effects; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D020298:U937 Cells
chemical_list: D001562:Benzimidazoles; D002232:Carbocyanines; D004791:Enzyme Inhibitors; D007653:Ketocholesterols; C007392:3,3'-dihexyl-2,2'-oxacarbocyanine; C008542:monodansylcadaverine; D002103:Cadaverine; C003001:7-ketocholesterol; C017807:bisbenzimide ethoxide trihydrochloride