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Using the tissue classifier add-on, the images of whole slides were segmented into tumor and stroma based on nuclear morphology. The whole image was segmented into four component images: 1) stroma, 2) tumor, 3) invasive margin, and 4) normal tissue. For each cell type, we recorded the percentage of cells and the mean of circularity ratio (Fig. 7).
Using the immune cell type as a model, we show in Fig. 8 the results of our image analysis. The cell nuclei were segmented by the tissue classifier add-on as shown in Fig. 8a. The nuclei were then analyzed by the spatial analysis module using several parameters.
The tissue classifier add-on is an adaptable plugin for the OpenSlide microscopic image analysis software that can generate a layer or two-dimensional map of the entire histological slide by segmenting cells and tumor regions. The tool (tissue classifier add-on) is based on the circularity and circularity ratio of the segmented nuclei. The segmented nuclei are presented as a single color in a binary image by the determination of the ratio of the major axis to the minor axis. The circularity ratio is a quantitative metric that measures the shape of a circular cell nucleus. The circularity ratio is defined as the ratio of the major axis to the minor axis. A nucleus with a circularity ratio of 1 is perfectly circular. With values approaching or reaching zero, the nucleus becomes increasingly elongated. An ideal cell shape is a perfect circle. The circularity ratio enables the determination of whether a nucleus is elliptical, circular, or elongated.
Cortical thickness increased significantly in both the femur and the tibia in the subchronic alcohol mice compared to the control mice. The BMD of subchronic alcohol mice at the end of the experiment was decreased significantly compared to controls in both femur and tibia. On the other hand, BV/TV in the subchronic alcohol mice were significantly decreased compared to the controls in both the femur and the tibia, while the percent bone marrow cavity was increased significantly in the subchronic alcohol mice compared to the controls in both femur and tibia (Table 2).
The values for relative bone volume and BMD were significantly decreased in mice from the group E. On the other hand, insignificant effects of subchronic alcohol consumption on relative bone volume without marrow cavity, bone surface and cortical bone thickness were observed. The results are summarized in Table 2. Representative reconstructed 3D images of the compact bone are illustrated in Fig. 2a, b. 827ec27edc