, Volume 35, Issue 1, pp 51–62 | Cite as

Factors influencing Giemsa band formation of human chromosomes

  • P. E. Crossen


Factors influencing a Giemsa banding method in which slides are treated with NaOH and then incubated in phosphate buffer were investigated. The study indicated that the removal of chromosomal proteins during fixation in acetic methanol is important for band formation. When fixatives containing formalin were used no banding occurred. Histones do not appear to be involved in band formation as neither of the two histone staining methods tested gave banding patterns. The age of the slide preparations was important, the best banding occurring on slides a week old. Romanovsky stains were the only stains to give banding, other stains resulted in distorted chromosomes. The composition of the incubation buffer had little effect on the quality of the banding. However liquid scintillation analysis of the phosphate buffer in which 3H-thymidine labelled preparations had been treated, revealed that thymidine is removed during incubation in buffer, and suggests that the degradation of thymidine is an important factor in band formation.


Phosphate Buffer Formalin Thymidine Banding Pattern Liquid Scintillation 
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  1. Alfert, M., Geschwind, I. I.: A selective staining method for the basic proteins of cell nuclei. Proc. nat. Acad. Sci. (Wash.) 39, 991–999 (1953).Google Scholar
  2. Ansede, A.: Human chromosome banding. Lancet 1972 I, 1184.Google Scholar
  3. Arrighi, F. E., Hsu, T. C.: Localization of heterochromatin in human chromosomes. Cytogenetics 10, 81–86 (1971).Google Scholar
  4. Baker, J. R.: Cytological technique. London: Metheun 1969.Google Scholar
  5. Bowbrow, M.: Cit. by Ridler, M. A. C., Banding patterns of metaphase chromosomes in Down's Syndrome. Lancet 1971 II, 354–356.Google Scholar
  6. Bowbrow, M., Madan, K., Pearson, P.: Staining of some specific regions of human chromosomes, particularly the secondary constriction of No. 9. Nature (Lond.) New Biol. 238, 122–124 (1972).Google Scholar
  7. Britten, R. J., Kohne, D. E.: Repeated sequences in DNA. Science 161, 529–540 (1968).Google Scholar
  8. Caspersson, T., Zech, L., Johansson, C.: Analysis of the human metaphase chromosome set by aid of DNA binding fluorescent agents. Exp. Cell Res. 62, 490–492 (1970).Google Scholar
  9. Crossen, P. E.: Giemsa banding patterns of human chromosomes. Clin. Genet. 3, 169–179 (1972).Google Scholar
  10. Crossen, P. E.: Human chromosome banding. Lancet 1972a II, 920–921.Google Scholar
  11. Cleaver, J. E.: Thymidine metabolism and cell kinetics. Amsterdam: North Holland 1967.Google Scholar
  12. Conn, H. J.: Biological stains (ed. R. D. Lillie). Baltimore: The Williams & Wilkins Co. 1969.Google Scholar
  13. Dick, C., Johns, E. W.: The effect of two acetic acid containing fixatives on the histone content of calf thymus deoxyribonucleoprotein and calf thymus tissue. Exp. Cell Res. 51, 626–632 (1968).Google Scholar
  14. Dutrillaux, B., Lejeune, J.: Sur une nouvelle technique d'analyse du caryotype humain. C. R. Acad. Sci. (Paris) 272, 2638–2640 (1971).Google Scholar
  15. Evans, H. J., Buckton, K. E., Sumner, A. T.: Cytological mapping of human chromosomes: Results obtained with quinacrine fluorescence and the acetic saline Giemsa techniques. Chromosoma (Berl.) 35, 310–325 (1971).Google Scholar
  16. Fand, S. B.: Environmental conditions for optimal Feulgen hydrolysis. Introduction to quantitative cytochemistry II (eds. G. L. Weid and G. F. Bahr), p. 209–221. London: Academic Press 1970.Google Scholar
  17. Ganner, E., Evans, H. J.: The relationship between patterns of DNA replication and of quinacrine fluorescence in the human chromosome complement. Chromosoma (Berl.) 35, 326–341 (1971).Google Scholar
  18. Grossman, L., Levine, S. S., Allison, W. S.: The reaction of formaldehyde with nucleotides and T. 2 bacteriophage DNA. J. molec. Biol. 3, 47–60 (1961).Google Scholar
  19. Hsu, T. C., Arrighi, F. E., Saunders, G. F.: Compositional heterogeneity of human heterochromatin. Proc. nat. Acad. Sci. (Wash.) 69, 1464–1466 (1972).Google Scholar
  20. Jones, K. W., Corneo, G.: Location of satellite and homogenous DNA sequences on human chromosomes. Nature (Lond.). New Biol 233, 268–271 (1971).Google Scholar
  21. Kato, H., Moriwaki, K.: Factors involved in the production of banded structures in mammalian chromosomes. Chromosoma (Berl.) 38, 105–120 (1972).Google Scholar
  22. Labelle, J. L., Brière, N.: Staining of nuclear basic proteins without deoxyribonucleic acid hydrolysis using heparin and alcian blue. Acta histochem. (Jena) 41, 338–348 (1971).Google Scholar
  23. Pardue, M. L., Gall, J. G.: Chromosomal localization of mouse satellite DNA. Science 168, 1356–1358 (1970).Google Scholar
  24. Saunders, G. F., Hsu, T. C., Getz, M. J., Simes, E. L., Arrighi, F. E.: Locations of human satellite DNA in human chromosomes. Nature (Lond.) New Biol. 236, 244–246 (1972).Google Scholar
  25. Schnedl, W.: Banding patterns of human chromosomes. Nature (Lond.) New Biol. 233, 93–94 (1971).Google Scholar
  26. Schnedl, W.: Analysis of the human karyotype using a reassociation technique. Chromosoma (Berl.). 34, 448–454 (1971a).Google Scholar
  27. Seabright, M.: A rapid banding technique for human chromosomes. Lancet 1971 II, 971–972.Google Scholar
  28. 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
  29. Subirana, J. A., Doty, P.: Kinetics of renaturation of denatured DNA. 1. Spectrophotometric results. Biopolymers 4, 171–187 (1966).Google Scholar
  30. 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
  31. Utakoji, T.: Differentiál staining patterns of human chromosomes treated with potassium permanganate. Nature (Lond.) New Biol. 239, 168–170 (1972).Google Scholar
  32. Wang, H. C., Federoff, S.: Banding of human chromosomes treated with trypsin. Nature (Lond.) New Biol. 235, 52–54 (1972).Google Scholar
  33. Weisblum, B., Haseth, P. L. de: Quinacrine a chromosome stain specific for deoxyadenylatedeoxythymidylate-rich regions in DNA. Proc. nat. Acad. Sci. (Wash.) 69, 629–632 (1972).Google Scholar
  34. Yosida, T. H., Sagai, T.: Banding pattern analysis of polymorphic karyotypes in the black rat by a new differential staining technique. Chromosoma (Berl.) 37, 387–394 (1972).Google Scholar

Copyright information

© Springer-Verlag 1973

Authors and Affiliations

  • P. E. Crossen
    • 1
  1. 1.Cytogenetics UnitChristchurch HospitalChristchurchNew Zealand

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