Skeletal muscle triad junction ultrastructure by Focused-Ion-Beam milling of muscle and Cryo-Electron Tomography

Basic and applied myology

Chyongere Hsieh¹  Michael Marko²  Terence Wagenknecht² 

[1] Department of Biomedical Sciences, School of Public Health, University at Albany, Albany, NY;New York State Department of Health, Wadsworth Cente, Empire State Plaza, Albany, NY

关键词: Cryo-electron microscopy;    Cryo-electron tomography;    Excitation-Contraction Coupling;    Focused-Ion-Beam milling;    Skeletal muscle;   

DOI  :  10.4081/ejtm.2015.4823

学科分类：农业科学（综合）

来源: Pagepress

【 摘 要 】

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.

【 授权许可】

CC BY