title: Repeatability, reproducibility and standardisation of a laser Doppler imaging technique for the evaluation of normal mouse hindlimb perfusion.
abstract: BACKGROUND
  Preclinical perfusion studies are useful for the improvement of diagnosis and therapy in dermatologic, cardiovascular and rheumatic human diseases. The Laser Doppler Perfusion Imaging (LDPI) technique has been used to evaluate superficial alterations of the skin microcirculation in surgically induced murine hindlimb ischemia. We assessed the reproducibility and the accuracy of LDPI acquisitions and identified several critical factors that could affect LDPI measurements in mice.
  
  
  METHODS
  Twenty mice were analysed. Statistical standardisation and a repeatability and reproducibility analysis were performed on mouse perfusion signals with respect to differences in body temperature, the presence or absence of hair, the type of anaesthesia used for LDPI measurements and the position of the mouse body.
  
  
  RESULTS
  We found excellent correlations among measurements made by the same operator (i.e., repeatability) under the same experimental conditions and by two different operators (i.e., reproducibility). A Bland-Altman analysis showed the absence of bias in repeatability (p = 0.29) or reproducibility (p = 0.89). The limits of agreement for repeatability were -0.357 and -0.033, and for reproducibility, they were -0.270 and 0.238. Significant differences in perfusion values were observed in different experimental groups.
  
  
  CONCLUSIONS
  Different experimental conditions must be considered as a starting point for the evaluation of new drugs and strategic therapies.
authors.affiliation: Dipartimento di Scienze Biomediche Avanzate, University Federico II, Via Pansini 5, 8013 Naples, Italy. adegreco@unina.it
authors.forename: Adelaide
authors.initials: A
authors.lastname: Greco
authors.forename: Monica
authors.initials: M
authors.lastname: Ragucci
authors.forename: Raffaele
authors.initials: R
authors.lastname: Liuzzi
authors.forename: Sara
authors.initials: S
authors.lastname: Gargiulo
authors.forename: Matteo
authors.initials: M
authors.lastname: Gramanzini
authors.forename: Anna Rita Daniela
authors.initials: AR
authors.lastname: Coda
authors.forename: Sandra
authors.initials: S
authors.lastname: Albanese
authors.forename: Marcello
authors.initials: M
authors.lastname: Mancini
authors.forename: Marco
authors.initials: M
authors.lastname: Salvatore
authors.forename: Arturo
authors.initials: A
authors.lastname: Brunetti
mesh_terms: D000818:Animals; D005260:Female; D006614:Hindlimb / Q000098:blood supply*; D006801:Humans; D017078:Laser-Doppler Flowmetry / Q000379:methods* / Q000592:standards*; D051379:Mice; D008833:Microcirculation / Q000502:physiology*; D010477:Perfusion / Q000379:methods* / Q000592:standards*; D012015:Reference Standards; D015203:Reproducibility of Results