The Sarcoplasmic Reticulum of Frog Slow and Twitch Muscle Fibers as Revealed by Stereoscopic High Voltage Electron Microscopy
Our present understanding of the distribution and morphology of the sarcoplasmic reticulum (SR) in frog slow and twitch muscle fibers has been derived largely from the examination of thin sections by electron microscopy. This conventional approach to the study of an organelle as complex as the SR is limited to a degree by section thickness, and the extraction of three-dimensional information must usually be gathered from an extensive collection of two-dimensional images. The present study represents an alternative approach to the problem of investigating the three-dimensional organization of the SR by utilizing high voltage electron microscopy (HVEM) and examining stereoscopic images of selectively stained 1.0 /μm thick slices of muscle tissue.Slow and twitch fibers from the distal fiber bundles of the frog (Rana pipiens) cruralis muscle were processed for electron microscopy according to the selective SR staining technique (DAB-H2O2 and Os-ferrocyanide) developed by Waugh. Tissue slices from 0.25 to 1.0 μm in thickness were cut on a diamond knife, mounted on grids either with or without plastic support films, and examined using the JEM-1000 microscope at the University of Colorado operating at an accelerating voltage of 1000 kV.