title: Improved Target Cell Selection and Counting Method for UroVysion Fluorescence in Situ Hybridization.
abstract: BACKGROUND
  The UroVysion Bladder Cancer Kit requires morphological analysis of 4', 6-diamino-2-phenylindole (DAPI)-stained nuclei to identify target cells for fluorescence in situ hybridization (FISH) signals. Reproducibility and efficiency of target cell selection and counting was evaluated by combining immunofluorescence staining of cytokeratin 7 (CK7) and proliferating cell nuclear antigen (PCNA) with DAPI staining.
  
  
  METHODS
  The reactivities to CK7, PCNA, and DAPI were compared between those for different ratios of T24 human bladder carcinoma cells and of cells from the urine of five healthy subjects. Two technicians independently performed five replicate cell counts of urine samples from four bladder cancer patients and one healthy subject.
  
  
  RESULTS
  The positive staining rates for CK7 and PCNA were similar to DAPI, but our method showed enhanced inter-observer repeatability and reduced operating time for signal counting.
  
  
  CONCLUSIONS
  Our proposed method showed better reproducibility and lesser operational time for signal counting than the DAPI method alone.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Hattori
authors.forename: Arizumi
authors.initials: A
authors.lastname: Kikuchi
authors.forename: Takahiro
authors.initials: T
authors.lastname: Sawamura
authors.forename: Osami
authors.initials: O
authors.lastname: Daimaru
authors.forename: Masanobu
authors.initials: M
authors.lastname: Horie
authors.forename: Takashi
authors.initials: T
authors.lastname: Deguchi
mesh_terms: D002277:Carcinoma / Q000175:diagnosis* / Q000652:urine; D045744:Cell Line, Tumor; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007211:Indoles; D053552:Keratin-7 / Q000032:analysis*; D018809:Proliferating Cell Nuclear Antigen / Q000032:analysis*; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis* / Q000652:urine
chemical_list: D007211:Indoles; D053552:Keratin-7; D018809:Proliferating Cell Nuclear Antigen; C007293:DAPI