Fixation effect of surepath preservative fluids using epidermal growth factor receptor mutation-specific antibodies for immunocytochemistry

Akihiko Kawahara, Tomoki Taira, Hideyuki Abe, Kosuke Watari, Yuichi Murakami, Chihiro Fukumitsu, Yorihiko Takase, Tomohiko Yamaguchi, Koichi Azuma, Jun Akiba, Mayumi Ono, Masayoshi Kage

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

BACKGROUND: Cytological diagnosis of respiratory disease has become important, not only for histological typing using immunocytochemistry (ICC) but also for molecular DNA analysis of cytological material. The aim of this study was to investigate the fixation effect of SurePath preservative fluids. METHODS: Human lung cancer PC9 and 11-18 cell lines, and lung adenocarcinoma cells in pleural effusion, were fixed in CytoRich Blue, CytoRich Red, 15% neutral-buffered formalin, and 95% ethanol, respectively. PC9 and 11-18 cell lines were examined by ICC with epidermal growth factor receptor (EGFR) mutation-specific antibodies, the EGFR mutation DNA assay, and fluorescence in situ hybridization. The effect of antigenic storage time was investigated in lung adenocarcinoma cells in pleural effusion by ICC using the lung cancer detection markers. RESULTS: PC9 and 11-18 cell lines in formalin-based fixatives showed strong staining of EGFR mutation-specific antibodies and lung cancer detection markers by ICC as compared with ethanol-based fixatives. DNA preservation with CytoRich Blue and CytoRich Red was superior to that achieved with 95% ethanol and 15% neutralbuffered formalin fixatives, whereas EGFR mutations by DNA assay and EGFR gene amplification by fluorescence in situ hybridization were successfully identified in all fixative samples. Although cytoplasmic antigens maintained high expression levels, expression levels in nuclear antigens fell as storage time increased. CONCLUSIONS: These results indicate that CytoRich Red is not only suitable for ICC with EGFR mutation-specific antibodies, but also for DNA analysis of cytological material, and is useful in molecular testing of lung cancer, for which various types of analyses will be needed in future.

Original languageEnglish
Pages (from-to)145-152
Number of pages8
JournalCancer Cytopathology
Volume122
Issue number2
DOIs
Publication statusPublished - Jan 1 2014

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Fingerprint Dive into the research topics of 'Fixation effect of surepath preservative fluids using epidermal growth factor receptor mutation-specific antibodies for immunocytochemistry'. Together they form a unique fingerprint.

  • Cite this

    Kawahara, A., Taira, T., Abe, H., Watari, K., Murakami, Y., Fukumitsu, C., Takase, Y., Yamaguchi, T., Azuma, K., Akiba, J., Ono, M., & Kage, M. (2014). Fixation effect of surepath preservative fluids using epidermal growth factor receptor mutation-specific antibodies for immunocytochemistry. Cancer Cytopathology, 122(2), 145-152. https://doi.org/10.1002/cncy.21355