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Silver‐enhanced colloidal‐gold labelling of rabbit kidney collecting‐duct cell surfaces imaged by scanning electron microscopy
Author(s) -
HERTER P.,
LAUBE G.,
GRONCZEWSKI J.,
MINUTH W. W.
Publication year - 1993
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1993.tb03364.x
Subject(s) - scanning electron microscope , colloidal gold , biotinylation , electron microscope , transmission electron microscopy , microscopy , biophysics , chemistry , labelling , materials science , pathology , biology , nanotechnology , optics , biochemistry , nanoparticle , medicine , physics , composite material
Summary 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‐μm‐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 (2000–4000 x) 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.