Silver-enhanced colloidal-gold labelling of rabbit kidney collecting-duct cell surfaces imaged by scanning electron microscopy.
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.
Herter
P
P
Max-Planck-Institut für Molekulare Physiologie, Dortmund, Germany.
Laube
G
G
Gronczewski
J
J
Minuth
W W
WW
D000375:Aging; D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004583:Electrons; D004848:Epithelium / Q000648:ultrastructure; D017979:Gold Colloid; D006651:Histocytochemistry / Q000379:methods; D007685:Kidney Tubules, Collecting / Q000648:ultrastructure*; D037102:Lectins; D008297:Male; D008855:Microscopy, Electron, Scanning; D019887:Peanut Agglutinin; D011817:Rabbits; D012542:Scattering, Radiation; D016622:Silver Staining; D013194:Staining and Labeling / Q000379:methods*
D017979:Gold Colloid; D037102:Lectins; D019887:Peanut Agglutinin