Premium
Optimization and validation of immunocytochemical detection of oestrogen receptors on cytospins prepared from fine needle aspiration (FNA) samples of breast cancer
Author(s) -
Srebotnik Kirbiš I.,
Us Krašovec M.,
Pogačnik A.,
Strojan Fležar M.
Publication year - 2015
Publication title -
cytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 48
eISSN - 1365-2303
pISSN - 0956-5507
DOI - 10.1111/cyt.12143
Subject(s) - immunostaining , concordance , kappa , medicine , immunohistochemistry , antigen retrieval , fine needle aspiration , breast cancer , pathology , cancer , antibody , nuclear medicine , biopsy , immunology , linguistics , philosophy
Objective To optimize and validate immunocytochemical ( ICC ) assessment of oestrogen receptors ( ER s) on cytospins prepared from fine needle aspiration ( FNA ) samples. Methods Optimal conditions and variability in ICC detection of ER s were established on cytospins prepared from the human breast cancer cell line MCF ‐7. Protocols that yielded adequate results were further validated on 52 FNA samples of resected breast cancer tumours using analysis of concordance with the ER status, determined by standard immunohistochemistry on corresponding formalin‐fixed, paraffin‐embedded tissue ( FFPET ). On 37 diagnostic FNA samples, manual immunostaining with antibody 1 D 5 was compared with automated immunostaining with antibody 6 F 11. Results The highest percentage of ER ‐positive MCF ‐7 cells with lowest variability was obtained on methanol‐fixed cytospins with or without microwave pre‐treatment: 72 ± 5% and 75 ± 7%, respectively. Microwave pre‐treatment was mandatory for P apanicolaou‐stained cytospins in order to achieve between 63 ± 14% and 67 ± 9% of ER ‐positive MCF ‐7 cells. The concordance between ICC assessment of ER s on FNA samples and corresponding FFPET sections was complete for methanol‐fixed cytospins (100%, kappa = 1) and adequate for P apanicolaou‐stained cytospins (94%, kappa = 0.84) and P apanicolaou‐stained smears (92%, kappa = 0.75). Complete agreement in ICC detection of ER s was obtained for manual immunostaining with antibody 1 D 5 and automated immunostaining with antibody 6 F 11. Conclusions Methanol ‐ fixed cytospins prepared from FNA samples ensure highly reliable ICC assessment of ER s, whereas P apanicolaou‐stained cytospins or smears are conditionally suitable because of the small risk of false negative results.