title: Scanning force microscopy of microtubules and polymorphic tubulin assemblies in air and in liquid.
abstract: We have investigated microtubules (MTs) and polymorphic assemblies, formed in vitro from isolated microtubule protein, by scanning force microscopy (SFM) in air and in liquid. Immobilization of MTs was achieved by placing a drop of the assembly solution on a polylysine-coated coverslip. After washing with taxol and air drying, the characteristic microtubular fibrous morphology appeared in the SFM. The MTs formed a network similar to that obtained by transmission electron microscopy (TEM). A height of approximately 9.5 nm for dried MTs was computed from the surface topography. Glutaraldehyde fixation of the MTs yielded higher structures (approximately 14 nm), which swelled to approximately 20 nm after rehydration, a value close to the MT diameter of approximately 25 nm determined from TEM images of ultrathin sections. The protofilament pattern of the MTs and surface attached MT-associated proteins were not apparent from SFM, although the height along the long axis of the MTs appeared slightly modulated. In addition to MTs, various polymorphic tubulin assemblies including ribbons, hoops and double-walled MTs were visualized by SFM.
authors.affiliation: Institut für Molekulare Biotechnologie, Forschungsgruppe Elektronenmikroskopie und Molekulare Zytologie, Jena, Germany.
authors.forename: W
authors.initials: W
authors.lastname: Vater
authors.forename: W
authors.initials: W
authors.lastname: Fritzsche
authors.forename: A
authors.initials: A
authors.lastname: Schaper
authors.forename: K J
authors.initials: KJ
authors.lastname: Böhm
authors.forename: E
authors.initials: E
authors.lastname: Unger
authors.forename: T M
authors.initials: TM
authors.lastname: Jovin
mesh_terms: D000388:Air; D000818:Animals; D001923:Brain Chemistry; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D018625:Microscopy, Atomic Force; D008854:Microscopy, Electron; D008870:Microtubules / Q000648:ultrastructure*; D011487:Protein Conformation; D012996:Solutions; D013552:Swine; D014404:Tubulin / Q000648:ultrastructure*
chemical_list: D046911:Macromolecular Substances; D012996:Solutions; D014404:Tubulin