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Immunocytochemical Reactivity Comparison between Formalin-Fixed and Liquid-Based Cytology-Fixed Specimens. 2023

Nanako Sakabe, and Sayumi Maruyama, and Chihiro Ito, and Yuka Shimoyama, and Tetsuya Komene, and Wataru Oboshi, and Shouichi Sato, and Kazuhisa Sudo, and Katsuhide Ikeda
Pathophysiology Sciences, Department of Integrated Health Sciences, Nagoya University Graduate School of Medicine, Nagoya, Japan, nanako@met.nagoya-u.ac.jp.

BACKGROUND Liquid-based cytology (LBC)-fixed samples can be used for preparing multiple specimens of the same quality and for immunocytochemistry (ICC); however, LBC fixing solutions affect immunoreactivity. Therefore, in this study, we examined the effect of LBC fixing solutions on immunoreactivity. METHODS Samples were cell lines, and specimens were prepared from cell blocks of 10% neutral buffered formalin (NBF)-fixed samples and the four types of LBC-fixed samples: PreservCyt®, CytoRich™ Red, CytoRich™ Blue, and TACAS™ Ruby, which were post-fixed with NBF. ICC was performed using 24 different antibodies, and immunocytochemically stained specimens were analyzed for the percentage of positive cells. RESULTS Immunoreactivity differed according to the type of antigen detected. For nuclear antigens, the highest percentage of positive cells of Ki-67, WT-1, ER, and p63 was observed in the NBF-fixed samples, and the highest percentage of positive cells of p53, TTF-1, and PgR was observed in the TACAS™ Ruby samples. For cytoplasmic antigens, the percentage of positive cells of CK5/6, Vimentin, and IMP3 in LBC-fixed samples was higher than or similar to that in NBF-fixed samples. The percentage of positive cells of CEA was significantly lower in CytoRich™ Red and CytoRich™ Blue samples than in the NBF-fixed sample (p < 0.01). Among the cell membrane antigens, the percentage of positive cells of Ber-EP4, CD10, and D2-40 was the highest in NBF-fixed samples and significantly lower in CytoRich™ Red and CytoRich™ Blue samples than that in NBF-fixed samples (p < 0.01). The NBF-fixed and LBC-fixed samples showed no significant differences in the percentage of positive cells of CA125 and EMA. CONCLUSIONS ICC using LBC-fixed samples showed the same immunoreactivity as NBF-fixed samples when performed on cell block specimens post-fixed with NBF. The percentage of positive cells increased or decreased based on the type of fixing solution depending on the amount of antigen in the cells. Further, the detection rate of ICC with LBC-fixed samples varied according to the type of antibody and the amount of antigen in the cells. Therefore, we propose that ICC using LBC-fixed samples, including detection methods, should be carefully performed.

UI MeSH Term Description Entries
D007150 Immunohistochemistry Histochemical localization of immunoreactive substances using labeled antibodies as reagents. Immunocytochemistry,Immunogold Techniques,Immunogold-Silver Techniques,Immunohistocytochemistry,Immunolabeling Techniques,Immunogold Technics,Immunogold-Silver Technics,Immunolabeling Technics,Immunogold Silver Technics,Immunogold Silver Techniques,Immunogold Technic,Immunogold Technique,Immunogold-Silver Technic,Immunogold-Silver Technique,Immunolabeling Technic,Immunolabeling Technique,Technic, Immunogold,Technic, Immunogold-Silver,Technic, Immunolabeling,Technics, Immunogold,Technics, Immunogold-Silver,Technics, Immunolabeling,Technique, Immunogold,Technique, Immunogold-Silver,Technique, Immunolabeling,Techniques, Immunogold,Techniques, Immunogold-Silver,Techniques, Immunolabeling
D003581 Cytodiagnosis Diagnosis of the type and, when feasible, the cause of a pathologic process by means of microscopic study of cells in an exudate or other form of body fluid. (Stedman, 26th ed) Cytodiagnoses
D005404 Fixatives Agents employed in the preparation of histologic or pathologic specimens for the purpose of maintaining the existing form and structure of all of the constituent elements. Great numbers of different agents are used; some are also decalcifying and hardening agents. They must quickly kill and coagulate living tissue. Fixative,Pickling Agents,Agents, Pickling
D005557 Formaldehyde A highly reactive aldehyde gas formed by oxidation or incomplete combustion of hydrocarbons. In solution, it has a wide range of uses: in the manufacture of resins and textiles, as a disinfectant, and as a laboratory fixative or preservative. Formaldehyde solution (formalin) is considered a hazardous compound, and its vapor toxic. (From Reynolds, Martindale The Extra Pharmacopoeia, 30th ed, p717) Formalin,Formol,Methanal,Oxomethane
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000093222 Cytology The study of single cell as a unit of living organisms. Cytopathology, a branch of this discipline, studies structural changes of cells in diseases and is used in diagnosing diseases. Cytopathology,Cytopathologies
D000906 Antibodies Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).
D000941 Antigens Substances that are recognized by the immune system and induce an immune reaction. Antigen

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