A new method of preparing embeddment-free sections for transmission electron microscopy: applications to the cytoskeletal framework and other three-dimensional networks.
Diethylene glycol distearate is used as a removable embedding medium to produce embeddment -free sections for transmission electron microscopy. The easily cut sections of this material float and form ribbons in a water-filled knife trough and exhibit interference colors that aid in the selection of sections of equal thickness. The images obtained with embeddment -free sections are compared with those from the more conventional epoxy-embedded sections, and illustrate that embedding medium can obscure important biological structures, especially protein filament networks. The embeddment -free section methodology is well suited for morphological studies of cytoskeletal preparations obtained by extraction of cells with nonionic detergent in cytoskeletal stabilizing medium. The embeddment -free section also serves to bridge the very different images afforded by embedded sections and unembedded whole mounts.
D000362:Agar; D003094:Collagen; D003599:Cytoskeleton / Q000648:ultrastructure*; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D008854:Microscopy, Electron / Q000379:methods*; D008938:Mitosis
D000362:Agar; D003094:Collagen