title: High-resolution multiphoton tomography and fluorescence lifetime imaging of UVB-induced cellular damage on cultured fibroblasts producing fibres.
abstract: BACKGROUND
  Multiphoton tomography (MPT) is suitable to perform both ex vivo and in vivo investigations of living skin and cell cultures with submicron resolution. Fluorescence lifetime imaging (FLIM) generates image contrast between different states of tissue characterized by various fluorescence decay rates. Our purpose was to combine MPT and FLIM to evaluate fibroblasts and collagen fibres produced in vitro.
  
  
  METHODS
  Fibroblast cultures, 2-4 days old, at a subconfluent stage, were evaluated before and after irradiation with a single UVB dose. One month old cultures stimulated with ascorbic acid were also assessed.
  
  
  RESULTS
  After UVB radiation, fibroblasts appear irregular in size, lose their alignment and show a decrease in fluorescence lifetime. One month-old fibroblasts, producing collagen fibres after stimulation with ascorbic acid, appear as small roundish structures intermingled by filaments showing a granular arrangement.
  
  
  CONCLUSIONS
  The combination of MPT and FLIM may be useful for the in vitro study of cell modifications induced by injurious or protective agents and drugs.
authors.affiliation: Department of Dermatology, University of Modena and Reggio Emilia, 41124 Modena, Italy.
authors.forename: Seidenari
authors.initials: S
authors.lastname: Stefania
authors.forename: Schianchi
authors.initials: S
authors.lastname: Simona
authors.forename: Azzoni
authors.initials: A
authors.lastname: Paola
authors.forename: Benassi
authors.initials: B
authors.lastname: Luisa
authors.forename: Borsari
authors.initials: B
authors.lastname: Stefania
authors.forename: Cautela
authors.initials: C
authors.lastname: Jennifer
authors.forename: Ferrari
authors.initials: F
authors.lastname: Chiara
authors.forename: French
authors.initials: F
authors.lastname: Paul
authors.forename: Giudice
authors.initials: G
authors.lastname: Stefania
authors.forename: Koenig
authors.initials: K
authors.lastname: Karsten
authors.forename: Magnoni
authors.initials: M
authors.lastname: Cristina
authors.forename: Talbot
authors.initials: T
authors.lastname: Clifford
authors.forename: Dunsby
authors.initials: D
authors.lastname: Christopher
mesh_terms: D017209:Apoptosis / Q000528:radiation effects; D002478:Cells, Cultured; D024022:Fibrillar Collagens / Q000528:radiation effects* / Q000648:ultrastructure*; D005347:Fibroblasts / Q000473:pathology* / Q000528:radiation effects*; D006801:Humans; D007089:Image Enhancement / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014466:Ultraviolet Rays / Q000009:adverse effects*
chemical_list: D024022:Fibrillar Collagens
keywords: collagen; extracellular; fibroblasts; fluorescence lifetime imaging; matrix; multiphoton tomography