Software based analyses of immunohistochemical staining are designed for obtaining quantitative, reproducible, and objective data. However, often times only a certain type of positive cells or structures need to be quantified thus whole image analysis cannot be performed. Such an example is Hofbauer placental cells, which show positivity of some antigens together with trophoblast, but only Hofbauer cells represent the regions of interest (ROIs). Two independent observers evaluated the immunohistochemical staining intensity of Hofbauer cells in placenta samples stained for cytoplasmic antigens by ImageJ, QuPath and light microscopy. Thus, the precise manual determination of ROIs, i.e. Hofbauer cells, was necessary. We detected low inter-observer variability in staining intensity. Almost perfect agreement between observers was reached for ImageJ and QuPath whilst substantial agreement was reached for light microscopy evaluation. As for the comparison of ImageJ, QuPath and light microscopy, the agreement of all three methods (identical immunohistochemical intensity) was achieved for 38.1% samples. The almost perfect agreement of staining intensities was reached between ImageJ and QuPath, and moderate agreement for comparison of the light microscopy to both software. Software analyses are much more time-consuming, thus their utilization is at least questionable to evaluate ROIs with selection.

基于软件的免疫组化染色分析旨在获得定量，可再现和客观的数据。但是，通常仅需要定量某种类型的阳性细胞或结构，因此无法执行整个图像分析。这样的例子是霍夫鲍尔胎盘细胞，它显示了某些抗原与滋养层的阳性，但是只有霍夫鲍尔细胞代表了目标区域（ROI）。两名独立的观察者通过ImageJ，QuPath和光学显微镜评估了胎盘样本中Hofbauer细胞的免疫组织化学染色强度，这些胎盘样本已被细胞质抗原染色。因此，需要精确手动确定ROI（即Hofbauer细胞）。我们发现观察者之间的染色强度差异较小。ImageJ和QuPath在观察者之间达成了近乎完美的协议，而在光学显微镜评估方面则达成了实质性协议。至于ImageJ，QuPath和光学显微镜的比较，对于38.1％的样品，所有三种方法（相同的免疫组织化学强度）均达到了一致。在ImageJ和QuPath之间达成了几乎完美的染色强度一致性，并且在将光学显微镜与这两种软件进行比较时达到了适度一致性。软件分析非常耗时，因此，其使用至少对于选择评估ROI值得怀疑。对于38.1％的样品，所有三种方法（相同的免疫组化强度）都达到了一致。在ImageJ和QuPath之间达成了几乎完美的染色强度一致性，并且在将光学显微镜与这两种软件进行比较时达到了适度一致性。软件分析非常耗时，因此，其使用至少对于选择评估ROI值得怀疑。对于38.1％的样品，所有三种方法（相同的免疫组化强度）都达到了一致。在ImageJ和QuPath之间达成了几乎完美的染色强度一致性，并且在将光学显微镜与这两种软件进行比较时达到了适度一致性。软件分析非常耗时，因此，其使用至少对于选择评估ROI值得怀疑。