Some factors affecting the preservation of the cytoskeleton of cells in culture for scanning electron microscopy.
This paper reports some preliminary findings on the effects of the composition of the detergent extraction medium on the morphological and molecular preservation of the cytoskeleton of Balb/c 3T3 cells as visualized by scanning electron microscopy and SDS-PAGE. Specific variables tested are the concentrations of sodium, potassium, calcium and magnesium ions, and the osmolarity of the medium. The results suggest that the ionic composition of the medium is more important than osmolarity in determining the composition of the cytoskeleton obtained with Triton X-100 detergent. Calcium is particularly deleterious to morphological preservation, and better preservation is obtained when potassium is the principal cation than when sodium is. The best preservation of cytoskeletal structures and the greatest molecular complexity was obtained by extracting in a buffer composed of 1mM EGTA, 4% polyethylene glycol 6000, 1 mM GTP and 100 mM PIPES, pH 6.9.
D000818:Animals; D002478:Cells, Cultured / Q000648:ultrastructure*; D002999:Clone Cells; D003902:Detergents; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D051379:Mice; D008807:Mice, Inbred BALB C; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D017830:Octoxynol; D011092:Polyethylene Glycols; D012964:Sodium; D013048:Specimen Handling / Q000379:methods; D013395:Sucrose
D003902:Detergents; D007202:Indicators and Reagents; D011092:Polyethylene Glycols; D013395:Sucrose; D017830:Octoxynol; D012964:Sodium