Pathophysiology Sciences, Department of Integrated Health Sciences, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Division of Clinical Laboratory, Chiba Prefectural Sawara Hospital, Katori, Japan.
Diagn Cytopathol. 2023 Sep;51(9):546-553. doi: 10.1002/dc.25178. Epub 2023 Jun 17.
Immunocytochemistry (ICC) is an indispensable technique to improve diagnostic accuracy. ICC using liquid-based cytology (LBC)-fixed specimens has been reported. However, problems may arise if the samples are not fixed appropriately. We investigated the relationship between the LBC fixing solution and ICC and the usefulness of antigen retrieval (AR) in LBC specimens.
Specimens were prepared from five types of LBC-fixed samples using cell lines and the SurePath™ method. ICC was performed using 13 antibodies and analyzed by counting the number of positive cells in the immunocytochemically stained specimens.
Insufficient reactivity was observed using ICC without heat-induced AR (HIAR) in nuclear antigens. The number of positive cells increased in ICC with HIAR. The percentage of positive cells was lower in CytoRich™ Blue samples for Ki-67 and in CytoRich™ Red and TACAS™ Ruby samples for estrogen receptor and p63 than in the other samples. For cytoplasmic antigens, the percentage of positive cells for no-HIAR treatment specimens was low in the three antibodies used. In cytokeratin 5/6, the number of positive cells increased in all LBC specimens with HIAR, and the percentage of positive cells in CytoRich™ Red and TACAS™ Ruby samples was significantly lower (p < .01). For cell membrane antigens, CytoRich™ Blue samples had a lower percentage of positive cells than the other LBC-fixed samples.
The combination of detected antigen, used cells, and fixing solution may have different effects on immunoreactivity. ICC using LBC specimens is a useful technique, but the staining conditions should be examined before performing ICC.
免疫细胞化学（ICC）是提高诊断准确性不可或缺的技术。已经有报道称使用液基细胞学（LBC）固定标本进行 ICC。然而，如果样本固定不当，可能会出现问题。我们研究了 LBC 固定液与 ICC 的关系以及 LBC 标本中抗原修复（AR）的有用性。
使用细胞系和 SurePath™ 方法从五种 LBC 固定样本制备标本。使用 13 种抗体进行 ICC，并通过计数免疫细胞化学染色标本中阳性细胞的数量进行分析。
在没有热诱导 AR（HIAR）的 ICC 中，核抗原的反应性不足。在具有 HIAR 的 ICC 中，阳性细胞的数量增加。在 Ki-67 的 CytoRich™ Blue 样本和雌激素受体和 p63 的 CytoRich™ Red 和 TACAS™ Ruby 样本中，阳性细胞的百分比低于其他样本。对于细胞质抗原，在三种抗体中，未经 HIAR 处理的标本的阳性细胞百分比较低。在细胞角蛋白 5/6 中，所有 LBC 标本的 HIAR 后阳性细胞数量增加，CytoRich™ Red 和 TACAS™ Ruby 样本的阳性细胞百分比明显较低（p<.01）。对于细胞膜抗原，CytoRich™ Blue 样本的阳性细胞百分比低于其他 LBC 固定样本。
检测抗原、使用的细胞和固定液的组合可能对免疫反应性有不同的影响。使用 LBC 标本进行 ICC 是一种有用的技术，但在进行 ICC 之前应检查染色条件。
