title: A critical analysis of three quantitative methods of assessment of hepatic steatosis in liver biopsies.
abstract: The issue of adequately quantitatively evaluating hepatic steatosis is still unresolved. Therefore, we compared three methods of quantitative assessment. Two groups of mice (n = 10 each) were fed standard chow (10% fat, SC group) or a high-fat diet (60% fat, HF group) for 16 weeks, and hepatic triglyceride (HT) and liver tissue were then studied. Paraplast-embedded tissues stained by hematoxylin and eosin (H-E) were compared to frozen sections stained by Oil Red-O (ORO). In addition, the volume density of steatosis (Vv[steatosis, liver]) was measured by point counting (P-C, sections H-E or ORO) or by image analysis (I-A, sections ORO). HT was significantly higher in the HF group (104% greater, P = 0.0004) than in the SC group. With P-C and H-E, Vv[steatosis, liver] was 4.80 ± 0.90% in the SC group and 33.50 ± 3.17% in the HF group (600% greater, P < 0.0001). With P-C and ORO, Vv[steatosis, liver] was 4.86 ± 0.89% in the SC group and 25.21 ± 1.27% in the HF group (420% greater, P < 0.0001). With I-A and ORO, Vv[steatosis, liver] was 4.17 ± 0.85% in the SC group and 23.35 ± 1.58% in the HF group (460% greater, P < 0.0001). Correlations between Vv[steatosis, liver] and HT were strong and significant in all methods. In conclusion, all methods were appropriate and reproducible. In P-C and H-E, there is a slight overestimation of steatosis in the HF animals in comparison to frozen sections and ORO; in frozen sections, differences between P-C and I-A are insignificant.
authors: Laboratory of Morphometry, Metabolism and Cardiovascular Diseases, Biomedical Center, State University of Rio de Janeiro, Av 28 de Setembro 87 fds, 20551-030 Rio de Janeiro, RJ, Brazil.
authors: Mariana
authors: M
authors: Catta-Preta
authors: Leonardo Souza
authors: LS
authors: Mendonca
authors: Julio
authors: J
authors: Fraulob-Aquino
authors: Marcia Barbosa
authors: MB
authors: Aguila
authors: Carlos Alberto
authors: CA
authors: Mandarim-de-Lacerda
mesh_terms: D000818:Animals; D001706:Biopsy; D005234:Fatty Liver / Q000473:pathology*; D016014:Linear Models; D008099:Liver / Q000737:chemistry / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D015588:Observer Variation; D014280:Triglycerides / Q000032:analysis
chemical_list: D014280:Triglycerides