Quality control of immunocytochemical staining of effusions using a standardized method of cell processing.
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.
Kuenen-Boumeester
V
V
Department of Pathology, Dr. Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
van Loenen
P
P
de Bruijn
E M
EM
Henzen-Logmans
S C
SC
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
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen