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 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.
D000818:Animals; D001202:Ascitic Fluid / Q000166:cytology*; D002021:Buffers; D002462:Cell Membrane / Q000648:ultrastructure; D002478:Cells, Cultured; D003584:Cytological Techniques; D003593:Cytoplasm / Q000648:ultrastructure; D005404:Fixatives; D005976:Glutaral; D051379:Mice; D009940:Organoids / Q000648:ultrastructure; D009993:Osmium Tetroxide; D013696:Temperature
D002021:Buffers; D005404:Fixatives; D009993:Osmium Tetroxide; D005976:Glutaral