title: Quality control of immunocytochemical staining of effusions using a standardized method of cell processing.
abstract: OBJECTIVE
  To improve the quality and reproducibility of immunocytochemical staining of effusions by using a standardized method of cell processing.
  
  
  METHODS
  The study included the specimens of 108 effusions (44 benign, 56 adenocarcinoma metastases and 8 mesotheliomas). Hemorrhagic effusions were lysed using isotonic ammonium chloride. All pellets were fixed in 1% paraformaldehyde dissolved in phosphate-buffered saline (PBS), pH 7.4, and washed in PBS. A Burker counting chamber was used to adjust the pellets to a standard cell concentration. The panel of monoclonal antibodies (MAbs) included MOC-31, Ber-EP4 and anticarcinoembryonic antigen (anti-CEA). The alkaline phosphatase/anti-alkaline phosphatase technique was applied.
  
  
  RESULTS
  Standardized material processing resulted in reproducible specimens with good preservation of cell morphology, reduction of nonspecific interaction and good immunostain intensity. MAbs MOC-31 and Ber-EP4 gave similar results: both were positive in all adenocarcinomas. Anti-CEA was positive in 73%. Benign effusions showed no expression. In contrast to the literature, seven mesotheliomas showed variable membranous expression of MOC-31 and Ber-EP4.
  
  
  CONCLUSIONS
  High-quality immunostaining results were obtained by using a standardized method of cell processing. MAbs MOC-31 and Ber-EP4 cannot be used as differentiation markers between mesotheliomas and adenocarcinomas. Discrepancies in immunocytochemical staining results may be caused partly by differences in cell preparation.
authors.affiliation: Department of Pathology, Dr. Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
authors.forename: V
authors.initials: V
authors.lastname: Kuenen-Boumeester
authors.forename: P
authors.initials: P
authors.lastname: van Loenen
authors.forename: E M
authors.initials: EM
authors.lastname: de Bruijn
authors.forename: S C
authors.initials: SC
authors.lastname: Henzen-Logmans
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis / Q000473:pathology / Q000556:secondary; D000911:Antibodies, Monoclonal; D001202:Ascitic Fluid / Q000737:chemistry / Q000473:pathology*; D001943:Breast Neoplasms / Q000175:diagnosis / Q000473:pathology / Q000556:secondary; D002272:Carcinoembryonic Antigen / Q000032:analysis / Q000276:immunology; D016889:Endometrial Neoplasms / Q000175:diagnosis / Q000473:pathology / Q000556:secondary; D004912:Erythrocytes / Q000473:pathology; D005260:Female; D005770:Gastrointestinal Neoplasms / Q000175:diagnosis / Q000473:pathology / Q000556:secondary; D006461:Hemolysis; D006801:Humans; D007150:Immunohistochemistry / Q000592:standards*; D007680:Kidney Neoplasms / Q000175:diagnosis / Q000473:pathology / Q000556:secondary; D008297:Male; D008654:Mesothelioma / Q000175:diagnosis / Q000473:pathology; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000473:pathology / Q000556:secondary; D010996:Pleural Effusion / Q000737:chemistry / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000175:diagnosis / Q000473:pathology / Q000556:secondary; D011786:Quality Control
chemical_list: D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen