Imaging Keratan Sulfate in Ocular Tissue Sections by Immunofluorescence Microscopy and LA-ICP-MS

ACS Appl Bio Mater. 2022 Feb 21;5(2):853-861. doi: 10.1021/acsabm.1c01240. Epub 2022 Jan 25.

Abstract

Carbohydrate-specific antibodies can serve as valuable tools to monitor alterations in the extracellular matrix resulting from pathologies. Here, the keratan sulfate-specific monoclonal antibody MZ15 was characterized in more detail by immunofluorescence microscopy as well as laser ablation ICP-MS using tissue cryosections and paraffin-embedded samples. Pretreatment with keratanase II prevented staining of samples and therefore demonstrated efficient enzymatic keratan sulfate degradation. Random fluorescent labeling and site-directed introduction of a metal cage into MZ15 were successful and allowed for a highly sensitive detection of the keratan sulfate landscape in the corneal stroma from rats and human tissue.

Keywords: ICP-MS; cornea; extracellular matrix; glycosaminoglycans; imaging; immunofluorescence; immunohistochemistry; keratan sulfate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Cornea / diagnostic imaging
  • Glycosaminoglycans*
  • Keratan Sulfate*
  • Microscopy, Fluorescence
  • Rats

Substances

  • Antibodies, Monoclonal
  • Glycosaminoglycans
  • Keratan Sulfate