Immunohistochemistry is primarily employed to visualize the localization of specific molecules in tissue samples. However, there is an increasing need for software-assisted quantitative assessment. In the present study, we performed inverted blue channel-based pseudoimmunofluorescence image analysis using original immunohistochemistry images. In human esophageal squamous cell carcinoma tissues, various humoral factors promote the phosphorylation of signaling proteins, including protein kinase B (Akt) and/or extracellular signal-regulated kinase 1/2 (ERK1/2), leading to tumor progression. Our method demonstrated applicability in the analysis of localized signaling proteins in histological sections. Relatively high phosphorylated Akt (p-Akt) intensity was observed in the cancer-stroma adjacent (Adj) and noncancerous regions of the superficial layer (SL). Furthermore, localized phosphorylated ERK1/2 (Thr202/Tyr204) was observed in the Adj of the SL and invasive front, distinct from the pattern of p-Akt (Ser473) and p-Akt (Thr308). In conclusion, pseudoimmunofluorescent immunohistochemistry image analysis is useful for the quantitative assessment and objective interpretation of localized signaling proteins in esophageal squamous cell carcinoma. The method can also be applied to analyze various immunohistochemistry images from diverse tissues.

中文翻译：

---

人食管鳞癌组织中磷酸化信号蛋白的伪免疫荧光免疫组化图像分析

免疫组织化学主要用于可视化组织样本中特定分子的定位。然而，对软件辅助定量评估的需求日益增长。在本研究中，我们使用原始免疫组织化学图像进行了基于倒置蓝色通道的伪免疫荧光图像分析。在人食管鳞状细胞癌组织中，多种体液因子促进信号蛋白的磷酸化，包括蛋白激酶 B (Akt) 和/或细胞外信号调节激酶 1/2 (ERK1/2)，从而导致肿瘤进展。我们的方法证明了在组织学切片中局部信号蛋白分析的适用性。在癌基质邻近 (Adj) 和浅层 (SL) 的非癌区域中观察到相对较高的磷酸化 Akt (p-Akt) 强度。此外，在 SL 和侵袭前沿的 Adj 中观察到局部磷酸化的 ERK1/2 (Thr202/Tyr204)，与 p-Akt (Ser473) 和 p-Akt (Thr308) 的模式不同。总之，伪免疫荧光免疫组织化学图像分析对于食管鳞状细胞癌中局部信号蛋白的定量评估和客观解释是有用的。该方法还可用于分析来自不同组织的各种免疫组织化学图像。