Location and identification of colloidal gold particles on the cell surface with a scanning electron microscope and energy dispersive analyzer.
The use of colloidal gold particles for locating cell surface components by scanning electron microscopy (SEM) has been restricted due to difficulties in the identification of these gold particles under SEM. It is shown here how the gold particles bound to cell surfaces can be located and identified under SEM using the secondary electron imaging (SEI) mode with an energy dispersive X-ray microanalyzer (EDS). This enables reliable identification of gold particles and good quality micrographs of the cells to be achieved at the same time. The distribution of receptors for two lectins, concanavalin A (ConA) and wheat germ agglutinin (WGA), on the surface of cultured Raji cells and human erythrocytes is presented as an example. Raji cells and erythrocytes were fixed with glutaraldehyde, post-fixed with a glutaraldehyde-tannic acid mixture and then incubated with ConA- or WGA-coated gold particles. After dehydration and critical point drying, the specimen filters were mounted on copper stubs and coated with carbon. The cells were examined on a JEOL TEMSCAN 100CX II electron microscope. The gold particles could be identified with the EDS analyzer, which was able to detect the Au spectrum when the electron beam was focused on single gold particles using a magnification of 100,000 or more. High-resolution photographs of the same cells were obtained up to the same magnification of 100,000.
Eskelinen
S
S
Department of Physiology, University of Oulu, Finland.
Peura
R
R
D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis*; D002478:Cells, Cultured; D003102:Colloids; D003208:Concanavalin A / Q000378:metabolism; D004577:Electron Probe Microanalysis; D004910:Erythrocyte Membrane / Q000032:analysis*; D005404:Fixatives; D006046:Gold / Q000032:analysis*; D006651:Histocytochemistry; D006801:Humans; D008855:Microscopy, Electron, Scanning; D011952:Receptors, Concanavalin A / Q000032:analysis*; D011975:Receptors, Mitogen / Q000032:analysis*; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D003102:Colloids; D005404:Fixatives; D011952:Receptors, Concanavalin A; D011975:Receptors, Mitogen; D014909:Wheat Germ Agglutinins; C027503:wheat germ agglutinin receptor; D003208:Concanavalin A; D006046:Gold