Skip to main content
Log in

Demonstration of chromosome replication by BrdU antibody technique and electron microscopy

  • Original Investigations
  • Published:
Human Genetics Aims and scope Submit manuscript

Summary

We describe the use of different reporter groups in visualizing replication patterns on metaphase chromosomes after BrdU incorporation by the BrdU antibody technique for the electron microscope. There is an inverse correlation between the density of the label and the size of the reporter particles. This observation alludes to stereo problems interfering with the access of the labeled antibodies into the chromatin. The use of silver enhancement enables easy detection of 1-nm and 5-nm gold particles, which make replication patterns visible in the electron microscope as does the diaminobenzidine/ H2O2 reaction. Possible consequences for the demonstration of replication patterns and for nonradioactive DNA in situ hybridization are discussed.

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

  • Barnicot NA, Huxley HE (1961) The electron microscopy of unsectioned human chromosomes. Hum Genet 25:253–258

    Google Scholar 

  • Burkholder GD, Wang HC (1978) Electron microscopy of differentially BrdU-substituted sister chromatids treated with sodium phosphate. Chromosoma 70:101–107

    Google Scholar 

  • Danscher G (1981) Localization of gold in biological tissue. A photochemical method for light and electron microscopy. Histochemistry 71:81–88

    Google Scholar 

  • Dolbeare F, Beisker W, Pallavicini MG, Vanderlaan M, Gray JW (1985) Cytochemistry for bromodeoxyuridine/DNA analysis: stoichiometry and sensitivity. Cytometry 6:521–530

    Google Scholar 

  • Dresser ME, Moses MJ (1979) Silver staining of synaptonemal complexes in surface spreads for light and electron microscopy. Exp Cell Res 121:416–419

    Google Scholar 

  • Drouin R, Messier PE, Richer CL (1988) Human chromosome banding specific for electron microscopy. Cytogenet Cell Genet 47:117–120

    Google Scholar 

  • Goodpasture C, Bloom SE (1975) Visualization of nucleolar organizer regions in mammalian chromosomes using silver staining. Chromosoma 53:37–50

    Google Scholar 

  • Goyanes VJ, Schvartzman JB (1983) Electron microscopy of sister chromatid exchanges. Cytogenet Cell Genet 36:612–616

    Google Scholar 

  • Messier PE, Jean P, Richer CL (1986) Easy transfer of selected mitoses from light to electron microscopy. Cytogenet Cell Genet 43:207–210

    Google Scholar 

  • Raap AK, Marijnen HGJ, Ploeg M van der (1986) Denaturation, renaturation and loss of DNA during in situ hybridization procedures. Acta Histochem (Jena) Suppl 32:243

    Google Scholar 

  • Ruzicka F (1971) Ein einfaches Verfahren zur Darstellung von humanen Metaphasenplatten für das Elektronenmikroskop. Humangenetik 13:199–204

    Google Scholar 

  • Ruzicka F, Schwarzacher HG (1974) The ultrastructure of human mitotic chromosomes and interphase nuclei treated by Giemsa banding techniques. Chromosoma 46:443–454

    Google Scholar 

  • Schwarzacher HG, Wolf U (1974) Methods in human cytogenetics. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Schwarzacher HG, Ruzicka F, Sperling K (1976) Electron microscopy of human banded and prematurely condensed chromosomes. In: Pearson PL, Lewis KR (eds) Chromosomes today, vol 5. Wiley, New York, pp 227–234

    Google Scholar 

  • Schwarzacher HG, Mikelsaar AV, Schnedel W (1978) The nature of the Ag-staining of nucleolus organizer regions. Cytogenet Cell Genet 20:24–39

    Google Scholar 

  • Speit G, Haupter S (1985) On the mechanism of differential Giemsa staining of bromodeoxyuridine-substituted chromosomes. II. Difference between the demonstration of sister chromatid differentiation and replication patterns. Hum Genet 70:126–129

    Google Scholar 

  • Taniguchi T, Takayama S (1987) Effects of bromodeoxyuridine substitution on metaphase chromosome structures examined by scanning electron microscopy. Chromosoma 95:13–16

    Google Scholar 

  • Vogel W, Schempp W, Puel V (1978) Silver-staining specificity in metaphases after incorporation of 5-bromodeoxyuridine (BUDR). Hum Genet 40:199–203

    Google Scholar 

  • Vogel W, Autenrieth M, Speit G (1986) Detection of bromodeoxyuridine-incorporation in mammalian chromosomes by a bromodeoxyuridine-antibody. Hum Genet 72:129–132

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vogel, W., Mehnert, K. & Pentz, S. Demonstration of chromosome replication by BrdU antibody technique and electron microscopy. Hum Genet 84, 237–240 (1990). https://doi.org/10.1007/BF00200566

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00200566

Keywords

Navigation