Skip to main content
Log in

Simple differential Giemsa staining of sister chromatids after treatment with photosensitive dyes and exposure to light and the mechanism of staining

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

The essential steps of the 33258 Hoechst-Giemsa method for differential chromatid staining consist of (1) 33258 Hoechst treatment, (2) exposure to light, and (3) Giemsa staining. The staining was shown to be a function of the concentration of 33258 Hoechst and the light exposure. The dye was successfully replaced by various metachromatic dyes such as thionine. Two simple methods are proposed. Failure of the pale stained chromatids to restore Giemsa affinity with urea and trypsin and the diminished Feulgen reaction after light exposure suggest that not masking proteins but photolysis of the BrdU-incorporated chromatid components in the presence of photosensitive dyes play a role in the differential staining.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Arrighi, E., Hsu, T. C.: Localization of heterochromatin in human chromosomes. Cytogenetics 10, 81–86 (1971)

    Google Scholar 

  • Bianchi, N. O.: Chromosomes of the rat. II. DNA replication sequence of bone marrow chromosomes in vivo. Cytologia (Tokyo) 31, 276–293 (1966)

    Google Scholar 

  • Bowen, E. J., Wokes, F.: Fluorescence of solutions. London: Longmans 1953

    Google Scholar 

  • Huang, C. C.: Induction of a high incidence of damage to the X chromosomes of Rattus (Mastomys) natalensis by base analogues, viruses, and carcinogens. Chromosoma (Berl.) 23, 162–179 (1967)

    Google Scholar 

  • Ikushima, T., Wolff, S.: Sister chromatid exchanges induced by light flashes to 5-bromodeoxyuridine- and 5-iododeoxyuridine substituted Chinese hamster chromosomes. Exp. Cell Res. 87, 15–19 (1974)

    Google Scholar 

  • Kato, H.: Spontaneous sister chromatid exchanges detected by a BUdR-labelling method. Nature (Lond.) 251, 70–72 (1974a)

    Google Scholar 

  • Kato, H.: Is isolabeling a false image ? Exp. Cell Res. 89, 416–420 (1974)

    Google Scholar 

  • Kim, A.: Chromatidaustausch und Heterochromatinveränderungen menschlicher Chromosomen nach BUdR-Markierung. Hum. Genet. 25, 179–188 (1974)

    Google Scholar 

  • Korenberg, J. R., Freedlender, E. F.: Giemsa technique for the detection of sister chromatid exchanges. Chromosoma (Berl.) 48, 355–360 (1974)

    Google Scholar 

  • Latt, S. A.: Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc. nat. Acad. Sci. (Wash.) 70, 3395–3399 (1973)

    Google Scholar 

  • Latt, S. A.: Sister chromatid exchanges, indices of human chromosome damage and repair: detection by fluorescence and induction by Mitomycin C. Proc. nat. Acad. Sci. (Wash.) 71, 3162–3166 (1974)

    Google Scholar 

  • Perry, P., Wolff, S.: New Giemsa method for the differential staining of sister chromatids. Nature (Lond.) 251, 156–158 (1974)

    Google Scholar 

  • Regan, J. D., Setlow, R. B., Ley, R. D.: Normal and defective repair of damaged DNA in human cells: a sensitive assay utilizing the photolysis of bromodeoxyuridine. Proc. nat. Acad. Sci. (Wash.) 68, 708–712 (1971)

    Google Scholar 

  • Seabright, M.: A rapid banding technique for human chromosomes. Lancet 1971 II, 971–972

    Google Scholar 

  • Shibatani, A., Naora, H.: Feulgen reaction and quantitative cytochemistry of desoxypentose nucleic acid. IV. Microspectrophotometric study of the Feulgen reaction in situ. Biochim. biophys. Acta (Amst.) 12, 515–521 (1953)

    Google Scholar 

  • Shiraishi, Y., Yosida, T. H.: Banding pattern analysis of human chromosomes by use of a urea treatment technique. Chromosoma (Berl.) 37, 75–83 (1972)

    Google Scholar 

  • Sugiyama, T.: The mechanism of specific staining of acid mucopolysaccharide by alcian blue: on the influence of urea treatment on the stainability. Acta Histochem. Cytochem. 1, 16–30 (1968)

    Google Scholar 

  • Sugiyama, T.: Specific vulnerability of the largest telocentric chromosome of rat bone marrow cells to 7,12-dimethylbenz(a)-anthracene. J. nat. Cancer Inst. 47, 1267–1275 (1971)

    Google Scholar 

  • Sugiyama, T.: Chromatid rearrangement and carcinogenesis. Gann Monogr. Cancer Res. (Tokyo) 17, 393–403 (1975)

    Google Scholar 

  • Sumner, A. T., Evans, H. J., Buckland, R. A.: New technique for distinguishing between human chromosomes. Nature (Lond.) New Biol. 232, 31–32 (1971)

    Google Scholar 

  • Wolff, S., Perry, P.: Differential Giemsa staining of sister chromatids and the study of sister chromatid exchanges without autoradiography. Chromosoma (Berl.) 48, 341–353 (1974)

    Google Scholar 

  • Zakharov, A. F., Egolina, N. A.: Differential spiralization along mammalian mitotic chromosomes. I. BUdR-reversed differentiation in Chinese hamster chromosomes. Chromosoma (Berl.) 38, 341–365 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Goto, K., Akematsu, T., Shimazu, H. et al. Simple differential Giemsa staining of sister chromatids after treatment with photosensitive dyes and exposure to light and the mechanism of staining. Chromosoma 53, 223–230 (1975). https://doi.org/10.1007/BF00329173

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00329173

Keywords

Navigation