A method for scanning electron microscopy of mitotic apparatus of cells in culture.
The mitotic apparatus of HeLa cells which had been covered with surface structures was successfully exposed for scanning electron microscopy. HeLa cells were cultured on carbon coated coverslips. After synchronization they were lysed with non-ionic detergent in a microtubule- stabilizingbuffer and embedded in styrene. Then the surface layer of the cells was sectioned away with a glass knife. Styrene was removed by treating the specimens with isoamylacetate and the cells were processed for scanning-electron microscopy. Various stages of mitotic apparatus were obtained with good preservation. Chromosomes appeared as aggregates of filamentous and granular structures. Microtubules appeared as filaments 40 nm in diameter.
D002877:Chromosomes, Human / Q000648:ultrastructure; D006367:HeLa Cells / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D008677:Metaphase; D008855:Microscopy, Electron, Scanning / Q000379:methods; D008938:Mitosis; D009685:Nuclear Envelope / Q000648:ultrastructure