DNA image-fluorimetry of individual human chromosomes

A microfluorimetric method has been developed for determination of DNA content in individual human chromosomes. The method is based on a preliminary identification of chromosomes with Hoechst 33258 followed by staining of the chromosomes with Feulgen reaction by using Schiff’s reagent type ethidium...

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Bibliographic Details
Published in:Cell and tissue biology 2013-07, Vol.7 (4), p.352-361
Main Authors: Agafonova, N. A., Sakuta, G. A., Rozanov, Yu. M., Shtein, G. I., Kudryavtsev, B. N.
Format: Article
Language:English
Subjects:
Biomedical and Life Sciences
Cell Biology
Life Sciences
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Summary:A microfluorimetric method has been developed for determination of DNA content in individual human chromosomes. The method is based on a preliminary identification of chromosomes with Hoechst 33258 followed by staining of the chromosomes with Feulgen reaction by using Schiff’s reagent type ethidium bromide-SO 2 and then by measuring the fluorescence intensity of the chromosomes by using an image analyzer. The method allows determining the DNA content of individual chromosomes with an accuracy up to 4.5 fg. The DNA content of individual human chromosomes and their p-and q-arms, as well as homologous chromosomes, were measured by using the developed method. It has been shown that the DNA content in chromosomes of the normal human karyotype is unstable and can fluctuate in some chromosomes within 35–40 fg.
ISSN:1990-519X
1990-5203
DOI:10.1134/S1990519X13040020