title: Automated topographical cell proliferation analysis.
abstract: BACKGROUND
  Cell proliferation is often studied using the incorporation of bromodeoxyuridine (BrdU). Immunohistochemical staining is then used to detect BrdU in the nucleus. To circumvent the observer bias and labor-intensive nature of manually counting BrdU-labeled nuclei, an automated topographical cell proliferation analysis method is developed.
  
  
  METHODS
  Sections stained with fluorescein-labeled anti-BrdU and counterstained with To-Pro-3 are scanned using confocal laser scanning microscopy (CLSM). For every point in the image, the nucleus density of BrdU-labeled nuclei and the total nucleus density of the neighborhood of that point are calculated from the BrdU and the To-Pro-3 signal, respectively. The ratio of these densities gives an indication of the amount of cell proliferation at that point. The automated measure is validated by comparing it with the ratio of BrdU-stained nuclei to the total number of nuclei obtained from a manual count.
  
  
  RESULTS
  A positive correlation is found between the automated measure and the ratios calculated from the manual counting (r = 0.86, P < 0.001). Calculating the topographical cell proliferation using the automated method is faster and does not suffer from interobserver variability.
  
  
  CONCLUSIONS
  Automated topographical cell proliferation analysis is a fast method to objectively find differences in cell proliferation within a tissue. This can be visualized by a topographical map that corresponds to the tissue under study.
authors.affiliation: Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht, The Netherlands.
authors.forename: E J
authors.initials: EJ
authors.lastname: van Raaij
authors.forename: D
authors.initials: D
authors.lastname: ten Berge
authors.forename: W
authors.initials: W
authors.lastname: Hage
authors.forename: A
authors.initials: A
authors.lastname: Brouwer
authors.forename: F
authors.initials: F
authors.lastname: Meijlink
authors.forename: J B
authors.initials: JB
authors.lastname: Maintz
authors.forename: F J
authors.initials: FJ
authors.lastname: Verbeek
mesh_terms: D000818:Animals; D001973:Bromodeoxyuridine / Q000032:analysis / Q000378:metabolism; D002455:Cell Division; D002467:Cell Nucleus / Q000737:chemistry* / Q000378:metabolism; D004622:Embryo, Mammalian / Q000737:chemistry / Q000378:metabolism; D005260:Female; D019044:Image Cytometry / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D008334:Mandible / Q000737:chemistry / Q000196:embryology / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D011247:Pregnancy; D015203:Reproducibility of Results
chemical_list: D001973:Bromodeoxyuridine