title: Quantitative histopathological analysis of cervical intra-epithelial neoplasia sections: methodological issues.
abstract: OBJECTIVE
  As part a Program Project to evaluate emerging optical technologies for cervical neoplasia, our group is performing quantitative histopathological analysis of biopsies from 1,800 patients. Several methodological issues have arisen with respect to this analysis: (1) Finding the most efficient way to compensate for staining intensity variation with out losing diagnostic information; (2) Assessing the inter- and intra-observer variability of the semi-interactive data collection; and (3) the use of non-overlapping cells from the intermediate layer only.
  
  
  METHODS
  Non-overlapping quantitatively stained nuclei were selected from 280 samples with histopathological characteristics of normal (199), koilocytosis (37), CIN 1 (18), CIN 2 (10) and CIN 3 (16). Linear discriminant analysis was used to assess the diagnostic information in three different feature sets to evaluate and compare staining intensity normalization methods. Selected feature values and summary scores were used to evaluate intra- and inter-observer variability.
  
  
  RESULTS
  The features normalized by the internal subset of the imaged cells had the same discriminatory power as those normalized by the control cells and by both normalization methods seem to have additional discriminatory power over the set of features which do not require normalization. The use of the internal subset decreased the image acquisition time by approximately 50% at each center, respectively. The intra- and inter-observer variability was of a similar size. Good performance was obtained by measuring the intermediate layer only.
  
  
  CONCLUSIONS
  The use of intensity normalization from a subset of the imaged non-overlapping intermediate layer cells works as well as or better than any of the other methods tested and provides a significant timesaving. Our intra- and inter-observer variability do not seem to affect the diagnostic power of the data. Although this must be tested in a larger data set, the use of intermediate layer cells only may be acceptable when using quantitative histopathology.
authors.affiliation: Dept. of Cancer Imaging, BC Cancer Agency, Vancouver, BC, Canada.
authors.forename: Martial
authors.initials: M
authors.lastname: Guillaud
authors.forename: Dennis
authors.initials: D
authors.lastname: Cox
authors.forename: Anais
authors.initials: A
authors.lastname: Malpica
authors.forename: Gregg
authors.initials: G
authors.lastname: Staerkel
authors.forename: Jasenka
authors.initials: J
authors.lastname: Matisic
authors.forename: Dirk
authors.initials: D
authors.lastname: Van Niekirk
authors.forename: Karen
authors.initials: K
authors.lastname: Adler-Storthz
authors.forename: Neal
authors.initials: N
authors.lastname: Poulin
authors.forename: Michele
authors.initials: M
authors.lastname: Follen
authors.forename: Calum
authors.initials: C
authors.lastname: MacAulay
mesh_terms: D002467:Cell Nucleus / Q000473:pathology; D048430:Cell Shape / Q000502:physiology; D002584:Cervix Uteri / Q000473:pathology*; D004847:Epithelial Cells / Q000473:pathology*; D005260:Female; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D016014:Linear Models; D008867:Microtomy / Q000379:methods / Q000592:standards; D015588:Observer Variation; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods / Q000592:standards; D002583:Uterine Cervical Neoplasms / Q000145:classification / Q000473:pathology*; D002578:Uterine Cervical Dysplasia / Q000145:classification / Q000473:pathology*