not-yet-known not-yet-known not-yet-known unknown 1 Pathological evaluation of steatosis 5-um thick sections were cut from the snap-frozen human liver specimens and placed on microscope slides. The sections were stained with ORO.30 ORO specifically stains TGs and cholesteryl oleate in the cytoplasm.31 ORO-staining operates well on frozen tissues, and it is commonly used for accurately identifying MiS and MaS.22,32–34 MaS typically appears as a single large lipid droplet in a liver cell, pushing the nucleus to the side and being at least as large as the nucleus. On the other hand, MiS was characterized by one or more small lipid droplets inside the cell’s cytoplasm that didn’t shift the nucleus and were smaller than it. Slides were digitized at 40X magnification in brightfield using an Aperio AT2 scanner (Leica Biosystems, Deer Park, IL, USA). In addition to the pathologist’s evaluation, an algorithm assessed the global HS by quantifying the ORO stain percentage based on the stained slide pixel area. As the algorithm mechanically summarizes the area of ORO-stained pixels (based on the area percentage), it cannot quantify MaS based on the Banff consensus (based on the cell percentage) with inability of recognizing boundary of cells. The difference of the area percentage and the cell percentage.12 1 Pathological evaluation of steatosis