title: Combined Ki-67 and Feulgen stain for morphometric determination of the Ki-67 labelling index.
abstract: In this note we present a combined Ki-67 and Feulgen stain for morphometric determination of the Ki-67 labelling index. The immunohistochemical part of this double staining technique is based on the alkaline-phosphatase-anti-alkaline-phosphatase (APAAP) method, visualizing the enzyme activity by the nitro-blue-tetrazolium chloride (NBT)/bromo-chloro-3-indolyl-phosphate (BCIP) technique. The NBT/BCIP complex resists the hydrolytic activity of the Feulgen stain. The staining method presented allows semi-automatic determination of both the total nucleus-area as well as the Ki-67 positive nucleus-area using a morphometric computer system. The Ki-67 labelling index thus achieved is based on the relative nuclear area of Ki-67 positive nuclei and is clearly more precise and efficient than the counting method using an ocular grid.
authors.affiliation: Department of Neuropathology, Medical School of the University of Saarland, Homburg, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Kolles
authors.forename: W
authors.initials: W
authors.lastname: Förderer
authors.forename: R
authors.initials: R
authors.lastname: Bock
authors.forename: W
authors.initials: W
authors.lastname: Feiden
mesh_terms: D000469:Alkaline Phosphatase / Q000276:immunology / Q000378:metabolism; D002467:Cell Nucleus / Q000648:ultrastructure; D004396:Coloring Agents; D005910:Glioma / Q000473:pathology; D006801:Humans; D006868:Hydrolysis; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D019394:Ki-67 Antigen; D009363:Neoplasm Proteins / Q000276:immunology / Q000378:metabolism*; D009423:Nervous System Neoplasms / Q000473:pathology; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D016612:Paraffin Embedding; D012394:Rosaniline Dyes; D013194:Staining and Labeling
chemical_list: D004396:Coloring Agents; C007722:Feulgen stain; D019394:Ki-67 Antigen; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D012394:Rosaniline Dyes; D000469:Alkaline Phosphatase