Silver-enhanced colloidal-gold labelling of rabbit kidney collecting-duct cell surfaces imaged by scanning electron microscopy

J Microsc. 1993 Aug;171(Pt 2):107-15. doi: 10.1111/j.1365-2818.1993.tb03364.x.

Abstract

The luminal cell surfaces of rabbit kidney cortical collecting-duct cells were labelled with peanut lectin (PNA) and investigated by scanning electron microscopy. Labelling was performed either on 20-microns-thick cryostat sections from prefixed and cryoprotected rabbit kidney tissue or on cultured collecting-duct epithelium using biotinylated PNA and a 6-nm colloidal-gold-coupled antibody against biotin. Colloidal-gold labels were detected at low magnification (2000-4000x) using silver enhancement. Coating with chromium allowed simultaneous imaging of both cell-surface morphology and labelling topography in the backscattered electron imaging mode. Our results show that PNA binding is specific for a subtype of intercalated cells equipped with microvilli on the luminal surface. The presented method promises to be useful for the identification of specific cell types in heterogeneous tissues.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aging
  • Animals
  • Animals, Newborn
  • Cells, Cultured
  • Electrons
  • Epithelium / ultrastructure
  • Gold Colloid
  • Histocytochemistry / methods
  • Kidney Tubules, Collecting / ultrastructure*
  • Lectins
  • Male
  • Microscopy, Electron, Scanning
  • Peanut Agglutinin
  • Rabbits
  • Scattering, Radiation
  • Silver Staining
  • Staining and Labeling / methods*

Substances

  • Gold Colloid
  • Lectins
  • Peanut Agglutinin