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, ors-collidine; and (c) the temperature of fixation. Results presented indicate that simultaneous fixation is far superior to sequential methods. Samples fixed sequentially at 4° C consistently had better morphological preservation than samples fixed under similar conditions at 23° C. With the exception ofs-collidine, which was totally unacceptable for in vitro in situ fixation on plastic, comparable results were noted with different buffer vehicles.
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Previous reports by Cohn and coworkers (1–3) have established that adherent peritoneal exudate cells (PEC) are monocytoid, i.e. macrophages. Thus, in this report, the term adherent peritoneal exudate cells will be used synonymously with macrophages.
Supported by a grant from the U.S. Veterans Administration entitled “A Comparative Study of Normal and Activated Macrophages.”
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Morris, R.E., Ciraolo, G.M., Cohen, D.A. et al. In situ fixation of cultured mouse peritoneal exudate cells: Comparison of fixation methods. In Vitro 16, 136–146 (1980). https://doi.org/10.1007/BF02831504
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DOI: https://doi.org/10.1007/BF02831504