Ferrocyanide enhancement of concanavalin A-ferritin and cationized ferritin staining blood cell surface glycoconjugates.
Ferrocyanide was used to enhance cationized ferritin and concanavalin A-ferritin (Con A-ferritin) staining of surface glycoconjugates of peripheral blood and bone marrow cells from rabbits and humans. The glutaraldehyde-fixed cells were stained with Con A-ferritin or cationized ferritin and then exposed to a ferrocyanide solution. The resulting cuboidal and irregular stain deposits averaged 50 nm in diameter when viewed with the transmission (TEM) and scanning electron microscope (SEM). Rabbit blood cells demonstrated more Con A binding sites than human blood cells and the decrease in binding sites observed with maturation of human granulocytic and erythrocytic cells was not evident in rabbit cells. Differences in binding of cationized ferritin to rabbit and human cell surfaces were less prominent than that observed for Con A. These results extend previous studies of blood cell surface glycoconjugates and demonstrate that ferrocyanide enhancement significantly facilitates SEM evaluation of Con A-ferritin and cationized ferritin bound to cell surfaces.
D000818:Animals; D002462:Cell Membrane / Q000032:analysis* / Q000648:ultrastructure; D003208:Concanavalin A; D004910:Erythrocyte Membrane / Q000032:analysis* / Q000648:ultrastructure; D004912:Erythrocytes / Q000032:analysis*; D005293:Ferritins; D005295:Ferrocyanides; D006023:Glycoproteins / Q000097:blood*; D006651:Histocytochemistry; D006801:Humans; D008214:Lymphocytes / Q000032:analysis*; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D011817:Rabbits
D005295:Ferrocyanides; D006023:Glycoproteins; D003208:Concanavalin A; D005293:Ferritins