title: Development of a computational high-throughput tool for the quantitative examination of dose-dependent histological features.
abstract: High-resolution digitalizing of histology slides facilitates the development of computational alternatives to manual quantitation of features of interest. We developed a MATLAB-based quantitative histological analysis tool (QuHAnT) for the high-throughput assessment of distinguishable histological features. QuHAnT validation was demonstrated by comparison with manual quantitation using liver sections from mice orally gavaged with sesame oil vehicle or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 0.001-30 μg/kg) every 4 days for 28 days, which elicits hepatic steatosis with mild fibrosis. A quality control module of QuHAnT reduced the number of quantifiable Oil Red O (ORO)-stained images from 3,123 to 2,756. Increased ORO staining was measured at 10 and 30 μg/kg TCDD with a high correlation between manual and computational volume densities (Vv ), although the dynamic range of QuHAnT was 10-fold greater. Additionally, QuHAnT determined the size of each ORO vacuole, which could not be accurately quantitated by visual examination or manual point counting. PicroSirius Red quantitation demonstrated superior collagen deposition detection due to the ability to consider all images within each section. QuHAnT dramatically reduced analysis time and facilitated the comprehensive assessment of features improving accuracy and sensitivity and represents a complementary tool for tissue/cellular features that are difficult and tedious to assess via subjective or semiquantitative methods.
authors.affiliation: Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, USA Center for Integrative Toxicology, Michigan State University, East Lansing, Michigan, USA.
authors.forename: Rance
authors.initials: R
authors.lastname: Nault
authors.affiliation: Institute for Cyber-Enabled Research, Michigan State University, East Lansing, Michigan, USA.
authors.forename: Dirk
authors.initials: D
authors.lastname: Colbry
authors.affiliation: Institute of Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Brandenberger
authors.affiliation: Center for Integrative Toxicology, Michigan State University, East Lansing, Michigan, USA Department of Pathobiology & Diagnostic Investigation, Michigan State University, East Lansing, Michigan, USA.
authors.forename: Jack R
authors.initials: JR
authors.lastname: Harkema
authors.affiliation: Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, USA Center for Integrative Toxicology, Michigan State University, East Lansing, Michigan, USA tzachare@msu.edu.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Zacharewski
mesh_terms: D000818:Animals; D001391:Azo Compounds; D003094:Collagen / Q000378:metabolism; D004396:Coloring Agents; D019295:Computational Biology / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D004785:Environmental Pollutants / Q000633:toxicity; D005234:Fatty Liver / Q000139:chemically induced / Q000473:pathology; D005260:Female; D005355:Fibrosis; D057166:High-Throughput Screening Assays / Q000379:methods*; D006652:Histological Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity; D014116:Toxicology / Q000379:methods*; D014617:Vacuoles / Q000473:pathology
chemical_list: D001391:Azo Compounds; D004396:Coloring Agents; D004785:Environmental Pollutants; D000072317:Polychlorinated Dibenzodioxins; D003094:Collagen; C011049:oil red O
keywords: TCDD; bioinformatics; hepatic; high throughput; histopathology; morphometry; quantitative