Quantification of Extracellular Matrix Fiber Systems Related to ADAMTS Proteins

Methods Mol Biol. 2020:2043:237-250. doi: 10.1007/978-1-4939-9698-8_19.

Abstract

ADAMTS (a disintegrin-like and metalloprotease domain with thrombospondin type 1 motifs) proteins regulate tissue homeostasis and extracellular matrix (ECM)-related pathogenesis. Some ADAMTS proteins interact with or process multiple ECM proteins, including fibrillins, fibronectin, and collagens. Therefore, characterization and quantification of these ECM fiber systems is essential to understand their functional relationship with ADAMTS proteins. Here we describe unbiased methods to quantify various aspects of ADAMTS-related ECM fiber systems in cell culture and in tissues. We focus on cell counting, overall fiber intensity, fiber length, and focal adhesion analysis in cell culture, and on the quantification of immunohistochemical and immunofluorescent tissue sections. We use ImageJ/Fiji, a widely used Java-based open source software which provides efficient and customizable quantification methods for microscopy images.

Keywords: Cell culture; Cell number; Extracellular matrix fibers; Fiji; Focal adhesions; ImageJ; Tissue sections.

MeSH terms

  • ADAMTS Proteins / metabolism*
  • Animals
  • Cell Count
  • Cells, Cultured
  • Extracellular Matrix / metabolism
  • Extracellular Matrix Proteins / metabolism*
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Mice
  • Software

Substances

  • Extracellular Matrix Proteins
  • ADAMTS Proteins