In situ fixation of cultured mouse peritoneal exudate cells: comparison of fixation methods

Mouse peritoneal exudate cells grown in vitro on plastic petri dishes were fixed in situ with both glutaraldehyde and osmium tetroxide by a variety of contemporary methods. The goal of the investigation was to determine which method resulted in the best ultrastructural preservation. The parameters b...

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Bibliographic Details
Published inIn vitro Vol. 16; no. 2; p. 136
Main Authors Morris, R E, Ciraolo, G M, Cohen, D A, Bubel, H C
Format Journal Article
LanguageEnglish
Published United States 01.02.1980
Subjects
Animals
Ascitic Fluid - cytology
Buffers
Cell Membrane - ultrastructure
Cells, Cultured
Cytological Techniques
Cytoplasm - ultrastructure
Fixatives
Glutaral
Mice
Organoids - ultrastructure
Osmium Tetroxide
Temperature
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Summary:Mouse peritoneal exudate cells grown in vitro on plastic petri dishes were fixed in situ with both glutaraldehyde and osmium tetroxide by a variety of contemporary methods. The goal of the investigation was to determine which method resulted in the best ultrastructural preservation. The parameters being tested included: (a) the method of fixation, i.e. either sequential or simultaneous; (b) the buffer vehicle for fixation, i.e. cocodylate, Mellonig's phosphate, Sorenson's phosphate, or s-collidine; and (c) the temperature of fixation. Results presented indicate that simultaneous fixation is far superior to sequential methods. Samples fixed sequentially at 4 degrees C consistently had better morphological preservation than samples fixed under similar conditions at 23 degrees C. With the exception of s-collidine, which was totally unacceptable for in vitro in situ fixation on plastic, comparable results were noted with different buffer vehicles.
ISSN:0073-5655
DOI:10.1007/BF02831504