Skip to main content
Log in

Ferrocyanide enhancement of concanavalin A-ferritin and cationized ferritin staining blood cell surface glycoconjugates

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Synopsis

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ackerman, G. A. (1975). Surface differentiation of hemopoietic cells demonstrated ultrastructurally with cationized ferritin.Cell Tiss. Res. 159, 23–37.

    Google Scholar 

  • Ackerman, G. A. &Freeman, W. H. (1978). Surface differentiation of developing human bone marrow cells demonstrated with concanavalin A-gold labeled horseradish peroxidase method.Anat. Rec. 190, 603.

    Google Scholar 

  • Ackerman, G. A. &Waksai, S. D. (1974). Ultrastructural localization of concanavalin A binding sites on the surface of differentiating hemopoietic cells.Cell Tiss. Res. 150, 331–42.

    Google Scholar 

  • Behnke, O. (1968). Electron microscopical observations of the surface coating of human blood platelets.Ultranstruct. Res. 24, 51–69.

    Google Scholar 

  • Bernhard, W. &Avrameas, S. (1971). Ultrastructural visualization of cellular carbohydrate components by means of concanavalin A.Exp. Cell Res. 64, 232–6.

    Google Scholar 

  • Danon, D., Goldstein, L., Marikovsky, Y. &Skutelsky, E. (1972). Use of cationized ferritin as a label of negative charges on cell surfaces.J. Ultrastruct. Res. 38, 500–10.

    Google Scholar 

  • Findlay, J. B. C. (1974). The receptor proteins for concanavalin A and lens culinaris phytohemagglutinin in the membrane of the human erythrocyte.J. biol. Chem. 249, 4398–403.

    Google Scholar 

  • Geoghegan, W. D. &Ackerman, G. A. (1977). Absorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A wheat germ agglutinin and goat antihuman immunoglobulin G on cell surfaces at the electron microscope level: a new method, theory and application.J. Histochem. Cytochem. 35, 1187–200.

    Google Scholar 

  • Leon, M. A. (1961). Concanavalin A reaction with human normal immunoglobulin G and myeloma immunoglobulin G.Science 158, 1325–6.

    Google Scholar 

  • Nicolson, G. L. &Singer, S. J. (1971). Ferritin-conjugated plant agglutinins as specific saccharide stains for electron microscopy: application to saccharides bound to cell membranes.Proc. Nat. Acad. Sci. (U.S.A.) 68, 942–5.

    Google Scholar 

  • Parmley, R. T., Martin, B. J. &Spicer, S. S. (1973). Staining of blood cell surfaces with a lectin-horseradish peroxidase method.J. Histochem. Cytochem. 21, 912–22.

    Google Scholar 

  • Parmley, R. T., Spicer, S. S. &Alvarez, C. J. (1978). Ultrastructural localization of non-heme cellular iron with ferrocyanide.J. Histochem. Cytochem. 26, 729–41.

    Google Scholar 

  • Perls, M. (1967). Nachweis von Eisenoxyd in gewissen Pigmenten.Virchows Arch. Path. Anat. 39, 42–8.

    Google Scholar 

  • Roth, J. &Binder, M. (1978). Colloidal gold, ferritin and peroxidase as markers for electron microscopic double labeling lectin techniques.J. Histochem. Cytochem. 26, 163–9.

    Google Scholar 

  • Skutelsky, E. &Danon, D. (1970). Electron microscopical analysis of surface charge labeling density at various stages of the erythroid line.J. Membrane Biol. 2, 173–9.

    Google Scholar 

  • Skutelsky, E., Danon, D., Wilchek, M. &Bayer, E. A. (1977). Localization of sialyl residues on cell surfaces by affinity cytochemistry.J. Ultrastruct. Res. 61. 325–35.

    Google Scholar 

  • Skutelsky, E. &Farquhar, M. G. (1976). Variations in distribution of Con A receptor sites and anionic groups during red blood cell differentiation in the rat.J. Cell Biol. 71, 218–31.

    Google Scholar 

  • Smith, S. B. &Revel, J. P. (1972). Mapping of concanavalin A binding sites on the surface of several cell types.Dev. Biol. 27, 434–41.

    Google Scholar 

  • Sumner, J. B. &Howell, S. F. (1936). The identification of the hemagglutinin of the Jack Bean with concanavalin AJ. Bacteriol. 34, 227–37.

    Google Scholar 

  • Femmink, J. H. M., Collard, J. G., Spits, H. &Roos, E. (1975). Comparative study of four cytochemical detection methods of concanavalin A binding sites on the cell membraneExp. Cell Res. 92, 307–22.

    Google Scholar 

  • Thiery, J. P. (1962). Etude des réactions cytochimique de fer au microscope électroniqueJ. Microscople 1, 127–36.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Parmley, R.T., Denys, F.R. & Alvarez, C.J. Ferrocyanide enhancement of concanavalin A-ferritin and cationized ferritin staining blood cell surface glycoconjugates. Histochem J 11, 379–389 (1979). https://doi.org/10.1007/BF01002766

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01002766

Keywords

Navigation