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Rapid multiple immunofluorescent staining for the simultaneous detection of cytokeratin and vimentin in the cytology of canine tumors
Author(s) -
Sawa Mariko,
Yabuki Akira,
Kohyama Moeko,
Miyoshi Noriaki,
Yamato Osamu
Publication year - 2018
Publication title -
veterinary clinical pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.537
H-Index - 51
eISSN - 1939-165X
pISSN - 0275-6382
DOI - 10.1111/vcp.12598
Subject(s) - pathology , cytokeratin , staining , vimentin , immunostaining , cytology , medicine , monoclonal antibody , immunohistochemistry , immunocytochemistry , antibody , immunology
Background Immunocytochemistry ( ICC ) is utilized as an advanced technique in veterinary cytology. In tumor diagnosis, cytokeratin and vimentin are markers used to distinguish the origin of tumor cells. Standard enzyme‐based ICC has limitations in clinical use; and therefore, more convenient and reliable methods are needed. Objectives The purpose of this study was to develop a rapid multiple immunofluorescent ( RMIF ) detection method for dual cytokeratin and vimentin staining on cytology slides in dogs. Methods Air‐dried smear samples from solid tumors and sediments of pleural effusions were prepared from dogs (n = 14) that were admitted to the Veterinary Teaching Hospital, Kagoshima University, Japan. Mouse monoclonal anti‐human cytokeratin ( AE 1/ AE 3) and rabbit monoclonal anti‐human vimentin ( SP 20) antibodies were used as primary antibodies, followed by staining with Alexa Fluor‐conjugated secondary antibodies. Staining using the RMIF method was compared with enzyme‐based ICC staining. Results Rapid multiple immunofluorescent immunostaining was clear and specific in the evaluated smears, whereas the enzyme‐based ICC showed nonspecific signals. By using the RMIF staining method, epithelial cells, mesenchymal cells, and mesothelial cells could be classified on a single smear of a pleural effusion. In smears of lymph nodes with epithelial tumor metastases, the RMIF method successfully detected metastatic epithelial tumor cells. Conclusions The RMIF method might be a useful tool for diagnostic cytology in veterinary medicine.