title: Quantification and Statistical Analysis Methods for Vessel Wall Components from Stained Images with Masson's Trichrome.
abstract: OBJECTIVE
  To develop a digital image processing method to quantify structural components (smooth muscle fibers and extracellular matrix) in the vessel wall stained with Masson's trichrome, and a statistical method suitable for small sample sizes to analyze the results previously obtained.
  
  
  METHODS
  The quantification method comprises two stages. The pre-processing stage improves tissue image appearance and the vessel wall area is delimited. In the feature extraction stage, the vessel wall components are segmented by grouping pixels with a similar color. The area of each component is calculated by normalizing the number of pixels of each group by the vessel wall area. Statistical analyses are implemented by permutation tests, based on resampling without replacement from the set of the observed data to obtain a sampling distribution of an estimator. The implementation can be parallelized on a multicore machine to reduce execution time.
  
  
  RESULTS
  The methods have been tested on 48 vessel wall samples of the internal saphenous vein stained with Masson's trichrome. The results show that the segmented areas are consistent with the perception of a team of doctors and demonstrate good correlation between the expert judgments and the measured parameters for evaluating vessel wall changes.
  
  
  CONCLUSIONS
  The proposed methodology offers a powerful tool to quantify some components of the vessel wall. It is more objective, sensitive and accurate than the biochemical and qualitative methods traditionally used. The permutation tests are suitable statistical techniques to analyze the numerical measurements obtained when the underlying assumptions of the other statistical techniques are not met.
authors.affiliation: IUMA Information and Communication Systems, University of Las Palmas de Gran Canaria, Campus Universitario de Tafira, Las Palmas de Gran Canaria, Spain.|Department of Telematic Engineering, University of Las Palmas de Gran Canaria, Campus Universitario de Tafira, Las Palmas de Gran Canaria, Spain.
authors.forename: Pablo
authors.initials: P
authors.lastname: Hernández-Morera
authors.affiliation: Department of Dermatology, Doctor Negrin University Hospital of Gran Canaria, Las Palmas de Gran Canaria, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Castaño-González
authors.affiliation: Institute for Technological Development and Innovation in Communications (IDeTIC), University of Las Palmas de Gran Canaria, Campus Universitario de Tafira, Las Palmas de Gran Canaria, Spain.
authors.forename: Carlos M
authors.initials: CM
authors.lastname: Travieso-González
authors.affiliation: Department of Morphology, University of Las Palmas de Gran Canaria, Campus de San Cristobal, Las Palmas de Gran Canaria, Spain.
authors.forename: Blanca
authors.initials: B
authors.lastname: Mompeó-Corredera
authors.affiliation: Department of Morphology, University of Las Palmas de Gran Canaria, Campus de San Cristobal, Las Palmas de Gran Canaria, Spain.|CliniVar, Clínica de Varices, Las Palmas de Gran Canaria, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Ortega-Santana
mesh_terms: D000465:Algorithms; D016000:Cluster Analysis; D003116:Color; D004396:Coloring Agents / Q000737:chemistry; D003287:Contrast Media; D001330:Electronic Data Processing; D005109:Extracellular Matrix / Q000473:pathology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009130:Muscle, Smooth / Q000473:pathology*; D015203:Reproducibility of Results; D012501:Saphenous Vein / Q000473:pathology*; D012984:Software; D013194:Staining and Labeling / Q000379:methods*; D014648:Varicose Veins / Q000473:pathology; D014689:Venous Insufficiency / Q000503:physiopathology
chemical_list: D004396:Coloring Agents; D003287:Contrast Media