Maciorowski Z, Veilleux C, Gibaud A, Bourgeois CA, Klijanienko J, Boenders J, Vielh P. 1997. Comparison of fixation procedures for fluorescent quantitation of DNA content using image cytometry. Cytometry 28: 123-129.
These authors evaluated four different fixatives for nuclear DNA quantitation of peripheral blood and breast cancer tumour cells, in the context of optimization of procedures for multi-colour, multi-parameter analyses, for example simultaneous DNA quantitation and FISH probe examination. The four treatments were “no fixative” (air-dried only), ethanol, ethanol / acetic acid, and paraformaldehyde / ethanol.
The fixation procedures were performed on cells already isolated from tissues, and already separated from other cells and substrates. Staining was with Propidium iodide, at concentrations comparable to current best practices recommendations, but the temperature and time of incubation seems to have varied for fixed vs. “no fixative” cells, and was never performed on ice.
Histograms of the relative DNA contents of various cells and treatments are presented, and none look completely useless. However, the authors state that the best treatment was with either paraformaldehyde or acetic acid with ethanol, depending on the other cellular parameters of interest.
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