Peculiarities of cytometrical methods of DNA content determination in the nucleus |
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| Authors: | Yu G Pichugin K A Semiyanov A V Chernyshev I G Palchikova L V Omelyanchyuk V P Maltsev |
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| Institution: | 1.Institute of Chemical Biology and Fundamental Medicine, Siberian Branch,Russian Academy of Sciences,Novosibirsk,Russia;2.Institute of Chemical Kinetics and Combustion, Siberian Branch,Russian Academy of Sciences,Novosibirsk,Russia;3.Novosibirsk University,Novosibirsk,Russia;4.Institute of Cytology and Genetics, Siberian Branch,Russian Academy of Sciences,Novosibirsk,Russia;5.Technological Design Institute of Scientific Instrument Engineering, Siberian Branch,Russian Academy of Sciences,Novosibirsk,Russia |
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| Abstract: | This work has proposed a new theoretical approach to analysis of histograms of DNA content, which are obtained by the method
of flow cytometry, in cells of Drosophila melanogaster imaginal discs. The precision of measurements of the DNA amount in G1 and G2(M) phases has been shown to be limited by precision of instrument tuning of zero of the flow cytometer. Use of the calculative
zero of the flow cytometer and of dividing cells as standards of the DNA content is able to increase severalfold the precision
of the DNA measurements in nuclei of the species. Comparative analysis of errors of various methods of measurement of the
DNA content in cell nuclei is also performed. For methods of flow fluorescent cytometry, confocal scanning, and cytophotometry
of the Feulgen-stained nuclei, it has been shown that, at present, the mean square errors of the DNA content measurements
are within the interval of values considered acceptable for biological studies (0.02 < CV < 0.06). |
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