Scanning Microscopy
Article Title
Conventional and High Resolution Scanning Electron Microscopy of Biological Sectioned Material
Abstract
Intracellular structures of embedded biological tissues (rat kidney, myocardium and small intestine) were observed by conventional-scanning electron microscopy (C-SEM) and high-resolution scanning electron microscopy (HR-SEM) after glass knife sectioning. C-SEM of semi -thin sections of material processed the same way as conventional transmission electron microscopy (TEM) provided strong backscattered electron (BSE)-dependent, two-dimensional secondary electron images (SEI(-)) which precisely integrated and further extended previous light microscopy (LM) observation of the same specimen. In addition, the three-dimensional (3-D) arrangement of intracellular organelles was appreciated using a mixture of acetone-soluble acrylic resin in place of epoxy resin embedding. Since the identification of such structures was hampered by the use of conventional fixations we introduced osmium maceration as a preliminary step to remove excess cytoplasmic matrix from the specimen. Consequently, semi-thin sections for LM and thin sections for TEM were obtained by sectioning of the tissue blocks . After resin removal, the sections were successfully observed in 3-D under a C-SEM. Finally, the deem bedded, osmium treated sections proved to be smooth enough to facilitate deposition of continuous, ultra-thin (1 nm) chromium films and, therefore , HR-SEM studies of macromolecular cell membrane structures.
Recommended Citation
Scala, C.; Cenacchi, G.; Preda, P.; Vici, M.; Apkarian, R. P.; and Pasquinelli, Gianandrea
(1990)
"Conventional and High Resolution Scanning Electron Microscopy of Biological Sectioned Material,"
Scanning Microscopy: Vol. 5
:
No.
1
, Article 13.
Available at:
https://digitalcommons.usu.edu/microscopy/vol5/iss1/13