title: Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography.
abstract: Cryo-electron tomography (cryo-ET) has emerged as perhaps the only practical technique for revealing nanometer-level three-dimensional structural details of subcellular macromolecular complexes in their native context, inside the cell. As currently practiced, the specimen should be 0.1- 0.2 microns in thickness to achieve optimal resolution. Thus, application of cryo-ET to intact frozen (vitreous) tissues, such as skeletal muscle, requires that they be sectioned. Cryo-ultramicrotomy is notoriously difficult and artifact-prone when applied to frozen cells and tissue, but a new technique, focused ion beam milling (cryo-FIB), shows great promise for "thinning" frozen biological specimens. Here we describe our initial results in applying cryo-FIB and cryo-ET to triad junctions of skeletal muscle.
authors.affiliation: ( ) New York State Department of Health, Wadsworth Cente, Empire State Plaza, Albany, NY ; ( ) Department of Biomedical Sciences, School of Public Health, University at Albany, Albany, NY , USA.
authors.forename: Terence
authors.initials: T
authors.lastname: Wagenknecht
authors.affiliation: ( ) New York State Department of Health, Wadsworth Cente, Empire State Plaza, Albany, NY.
authors.forename: Chyongere
authors.initials: C
authors.lastname: Hsieh
authors.affiliation: ( ) New York State Department of Health, Wadsworth Cente, Empire State Plaza, Albany, NY.
authors.forename: Michael
authors.initials: M
authors.lastname: Marko
keywords: Cryo-electron microscopy; Cryo-electron tomography; Excitation-Contraction Coupling; Focused-Ion-Beam milling; Skeletal muscle