Introduction/Objective: Liquid-based cytology (LBC) is advantageous as multiple stained specimens can be prepared and used for additional assays such as immunocytochemical and molecular-pathological investigations. Two types of preservative-fixative solutions (fixatives) are used for nongynecologic specimens used in the BD SurePath-LBC (SP-LBC) method, and their components vary. However, few studies have evaluated the differences in antigen-retaining ability between these fixatives. Therefore, we investigated and compared the antigen-retaining ability of the fixatives in immunocytochemical staining (ICC) under long-term storage conditions. Materials and Methods: Sediments of cultured RAJI cells (derived from Burkitt’s lymphoma) were added to each fixative (red and blue) and stored at room temperature for a specified period (1 h; 1 week; and 1, 3, and 6 months). The specimens were then prepared using the SP-LBC method and subjected to ICC. Positivity rate was calculated using the specimens fixed at room temperature for 1 h as a control. Antibodies against Ki67 expressed in the nucleus and against CD20 and leukocyte common antigen (LCA) expressed on the cell membrane were used. Results: For CD20 and LCA, the positivity rate increased with time in the red fixative compared with that in the control. In the blue fixative, the positivity rate was highest at 1 h and was maintained at a high level throughout the storage period. In contrast, the Ki67 positivity rate was highest at 1 h in both red and blue fixatives and markedly decreased with time. Therefore, although refrigerated (8°C) storage was used, no improvement was noted. Conclusions: Long-term storage is possible for cell membrane antigens at room temperature; however, it is unsuitable for intranuclear antigens. Therefore, we conclude that suitable fixative type and storage temperature differ based on antigen location. Further investigation is warranted.
Acta Cytologica

中文翻译：

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液基细胞学固定液的抗原性保留能力的功效：长期储存的免疫细胞化学

介绍/目的：液基细胞学 (LBC) 具有优势，因为可以制备多个染色标本并用于其他检测，例如免疫细胞化学和分子病理学研究。BD SurePath-LBC (SP-LBC) 方法中使用的非妇科标本有两种类型的防腐剂-固定液（固定液），它们的成分各不相同。然而，很少有研究评估这些固定剂之间抗原保留能力的差异。因此，我们研究并比较了长期储存条件下固定剂在免疫细胞化学染色 (ICC) 中的抗原保留能力。材料和方法：将培养的 RAJI 细胞（源自伯基特淋巴瘤）的沉淀物加入每种固定剂（红色和蓝色）中，并在室温下储存指定的时间（1 小时；1 周；以及 1、3 和 6 个月）。然后使用 SP-LBC 方法制备样品并进行 ICC。使用在室温下固定 1 小时的样品作为对照计算阳性率。使用了针对细胞核中表达的 Ki67 以及针对细胞膜上表达的 CD20 和白细胞共同抗原 (LCA) 的抗体。结果：对于 CD20 和 LCA，与对照相比，红色固定液的阳性率随时间增加。在蓝色固定液中，阳性率在 1 h 时最高，并在整个储存期间保持在高水平。相比之下，红色和蓝色固定剂的 Ki67 阳性率在 1 小时时最高，并随时间显着降低。因此，尽管使用了冷藏 (8°C) 储存，但没有发现任何改善。结论：细胞膜抗原可在室温下长期保存；然而，它不适用于核内抗原。因此，我们得出结论，合适的固定剂类型和储存温度因抗原位置而异。有必要进一步调查。
细胞学学报