Stepwise Protocol for Cytospin-enhanced Smearing for Scraped Corneal Epithelial Cells

Appl Immunohistochem Mol Morphol. 2016 Jan;24(1):71-3. doi: 10.1097/PAI.0000000000000168.

Abstract

Proteins and antigens present on the cell surface are usually determined by immunofluorescence staining. Uniform distribution of cells is required to appreciate the presence of surface proteins. Improper smearing or crushing of the corneal epithelial cells can potentially destroy the cellular integrity. Thus a simplified, systemic method was designed to smear the cells scraped from the cornea. The procedure includes trypsinisation for dissociation of corneal epithelial cells and cytospinning for concentrating the cells in a smear. The standardized protocol was found to be efficient in maintaining the integrity of the corneal epithelial cells and also the distribution of the cells in the smear.

MeSH terms

  • Biomarkers / metabolism
  • Cell Adhesion
  • Cell Separation / methods*
  • Centrifugation
  • Cornea / cytology*
  • Cornea / metabolism
  • Epithelial Cells / cytology*
  • Epithelial Cells / metabolism
  • Fluorescent Antibody Technique
  • Gene Expression
  • Humans
  • Microscopy, Fluorescence
  • Receptors, TNF-Related Apoptosis-Inducing Ligand / genetics*
  • Trypsin / chemistry

Substances

  • Biomarkers
  • Receptors, TNF-Related Apoptosis-Inducing Ligand
  • TNFRSF10A protein, human
  • Trypsin