Open AccessSkeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography
Author(s) -
Terence Wagenknecht,
Chyongere Hsieh,
Michael Marko
Publication year - 2015
Publication title -
european journal of translational myology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.271
H-Index - 6
eISSN - 2037-7460
pISSN - 2037-7452
DOI - 10.4081/ejtm.2015.4823
Subject(s) - cryo electron tomography , focused ion beam , triad (sociology) , context (archaeology) , materials science , skeletal muscle , electron microscope , cryo electron microscopy , electron tomography , ultrastructure , biophysics , biomedical engineering , tomography , nanotechnology , anatomy , ion , chemistry , transmission electron microscopy , biology , optics , scanning transmission electron microscopy , physics , medicine , psychology , paleontology , organic chemistry , psychoanalysis
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.