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Simultaneous fixation using glutaraldehyde and osmium tetroxide or potassium ferricyanide‐reduced osmium for the preservation of monogenean flatworms: An assessment for Merizocotyle icopae
Author(s) -
Cribb Bronwen,
Armstrong Wendy,
Whittington Ian
Publication year - 2004
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.20015
Subject(s) - glutaraldehyde , osmium tetroxide , osmium , fixation (population genetics) , potassium ferricyanide , chemistry , pipette , ferricyanide , fixative , chromatography , electron microscope , biochemistry , inorganic chemistry , physics , cytoplasm , ruthenium , optics , gene , catalysis
Simultaneous fixation was investigated for a marine organism: the monogenean flatworm ectoparasite Merizocotyle icopae . Four protocols for primary fixation were compared: 3% glutaraldehyde alone in 0.1M cacodylate buffer for a minimum of 2 hours; 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, until tissues darkened (5–20 minutes); 1% glutaraldehyde in 0.1M cacodylate buffer in combination with 0.5% potassium ferricyanide‐reduced osmium until tissues darkened (5–20 minutes); 1% glutaraldehyde in combination with 1% osmium tetroxide, both in 0.1M cacodylate buffer, for 30 minutes. The study confirms that the standard method for transmission electron microscopic fixation (first listed protocol) routinely applied to platyhelminths is optimal for ultrastructural preservation, but some simultaneous fixation methods (second and third listed protocols) are acceptable when rapid immobilization is needed. Scanning electron microscopic preparations may be improved using simultaneous primary fixation. Microsc. Res. Tech. 63:102–110, 2004. © 2004 Wiley‐Liss, Inc.