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Improved ultrastructural preservation of rat ciliary body after high pressure freezing and freeze substitution: A perspective view based upon comparison with tissue processed according to a conventional protocol or by osmium tetroxide/microwave fixation
Author(s) -
Eggli Peter S.,
Graber Werner
Publication year - 1994
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.1070290103
Subject(s) - osmium tetroxide , ultrastructure , fixation (population genetics) , fibrocyte , anatomy , cryofixation , biophysics , chemistry , biology , microbiology and biotechnology , biomedical engineering , electron microscope , biochemistry , medicine , physics , gene , optics
Conventional fixation of the delicate, highly folded rat ciliary body and its iridial extension, as well as of vitreal structures, is associated with the induction of a number of artifacts, thus limiting the reliability of morphological interpretations. Improved ultrastructural preservation may be achieved by microwave heating in combination with osmium tetroxide fixation. This protocol, although simple and cheap, yields results, particularly with respect to the extracellular matrix compartment between inner and outer ciliary epithelial cells, which are not greatly inferior to those obtained by implementing the sophisticated high pressure freezing and freeze substitution technique. The latter affords good to very good ultrastructural preservation of epithelium and stromal components, such as blood vessels, neural elements, smooth muscle cells, fibrocytes, and free cells, up to a depth of 50–100 μm from the tissue surface. Its superiority over osmium tetroxide/microwave fixation is revealed in the cytoplasmic, intraorganellar, and vitreal matrix compartments, which incur no obvious losses. © 1994 Wiley‐Liss, Inc.