![]() ![]() All these scoring systems use different categories for the proportion of tissues or cells stained. H-score, Allred-score, and Immunoreactive score are considered as a “gold standard” of combined scoring system in IHC data evaluation and presentation. The overall score of the staining intensity typically has four categories: negative (0), weak (1), moderate (2), and strong (3). The existing clinical scoring process is based on two characteristics: overall staining intensity and the proportion of tissue or cells stained. Semi-quantitative scoring systems are widely used to convert subject perception of IHC-marker expression into (semi)quantitative data, which is then used for statistical analyses and establishing of the conclusions. The stained sample slides are generally evaluated under the light microscope by trained pathologist or researcher. The visualization of the antibody-antigen reaction is accomplished by using the relevant chromogen. Antigens of interest in formalin-fixed paraffin-embedded tissues are detected by specific antibodies. Immunohistochemistry (IHC) is an effective, well-established and widely accepted method for localizing the expression of a specific proteins in tissues and it is used in both clinical and research practice. ![]() Software analyses are much more time-consuming, thus their utilization is at least questionable to evaluate ROIs with selection. 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. 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. Almost perfect agreement between observers was reached for ImageJ and QuPath whilst substantial agreement was reached for light microscopy evaluation. We detected low inter-observer variability in staining intensity. Thus, the precise manual determination of ROIs, i.e. Two independent observers evaluated the immunohistochemical staining intensity of Hofbauer cells in placenta samples stained for cytoplasmic antigens by ImageJ, QuPath and light microscopy. 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). However, often times only a certain type of positive cells or structures need to be quantified thus whole image analysis cannot be performed. Software based analyses of immunohistochemical staining are designed for obtaining quantitative, reproducible, and objective data. ![]()
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