Fig. 3.

Analysis of infiltrating cells (lymphocytes) using Image-Pro Plus. (A) Left image: original image of tissue specimen. Right image: analyzed image of infiltration cells. Green-colored objects were infiltrating cells, which were recognized using a size limitation of 5–45 μm². The area of infiltrating cells was measured. Light blue-colored objects represent nuclei. By adding measurement conditions with fill holes, area inside the contour of a nucleus was filled, and the nucleus was recognized as one object, whose size was larger than 45 μm². Clustered infiltrating cells (red arrows) had sizes larger than 45 μm², so they were not recognized as infiltrating cells in this experiment. Bar = 20 μm. (B) Total area of infiltrating cells in the liver tissue with various grades of inflammatory cell infiltration. For analysis, JPEG images of 4 mm² areas at 5 points were extracted from the whole slide image. (C) Representative images of various grades (left, center, and right images: normal, −; slight, +; and moderate, ++, respectively) of cell infiltration. Bar = 100 μm.