The progress of technology within the Cytogenetics Lab


  • 1955. The 22 nd of December Mr. Tjio at the University of Lund could observe chromosomes much better spread and bigger compared to the ones usually obtained; it was definitely clarified and became universally accepted that the number of chromosomes for the human species is 46.
  • 1959: a small extra-numerary chromosome (21) is detected by Lejeune, who was observing the caryotypes of nine patients suffering for the Down Syndrome.
    In the same year, Jacobs discovers that in Klinefelter’s Syndrome male patients a second “X” chromosome is present.
  • 1960-1970: Moorehead starts to use stimulated lymphocytes for post-natal diagnosis. Several banding techniques are introduced in these years by Caspersson (QFQ), Seabright (GTG), Dutrillaux (High Resolution).
  • 1980: a new medium, specially formulated by Chang to significantly decrease the culturing time of amniocytes, becomes commercially available, thus enhancing the pre-natal diagnostic procedures.
  • 1983: On February 3rd in the Cytogenetic Laboratory of the Ist Clinica Ostetrico-Ginecologica “Mangiagalli” (University of Milan), a trisomy 21 was diagnosed from cells in spontaneous mitosis obtained from a sample of Chorionic Villi ( CVS) taken during the first trimester of pregnancy (11).
  • 1990: starting from the end of this decade, the image analysis systems allow a growing number of cytogenetics laboratories to perform the caryograms quicker and easier.
  • 2002: Claussen finally shows that the effect of the hypotonic solution on the swelling of the mitotic cells during evaporation of the fixative is, in reality, ‘based on an interaction between acetic acid, water and cell proteins’.
  • 2006: On occasion of the IX Congress of the Italian Society of Human Genetics (SIGU), G. Terzoli and M. Di Segni proposed a possible mechanism to explain the swelling of both cytoplasmatic and chromosome proteins. According to their hypothesis, it is the protein hydration that produces, respectively, the spreading and the banding of the chromosomes
  • 2013: At the end of this year we had the publication of the study of lymphocytes during each stage of the chromosome preparation protocol by Fourier transform infrared (FTIR) spectroscopy; this study provided evidence at molecular level of the crucial role of the bound water to the cytoplasmic matrix and to the chromatin in determining the chromosome spreading and swelling. The FTIR results underlined the need to perform the last fixative-evaporation stage in standardized and optimized temperature and relative humidity conditions.

  • Bibliography

    1) Tjio, J.H., Levan, A.: The chromosome number of man. Hereditas 42, 1-6 (1956).
    2) Lejeune J., Gautier M., Turpin R.: Etude des chromosomes somatiques de neuf enfants mongoliens. C.R. Acad. Sci. Paris 248, 1721-1722, (1959).
    3) Jacobs P.A., Strong J.A.: A case of human intersexuality having a possible XXY sex determining mechanism. Nature 183, 302-303, (1959).
    4) Moorhead P.S., Nowell P.C., Mellman W.J. et al.: Chromosome preparations of leucocytes cultured from human peripheral blood. Exp. Cell. Res. 20, 613-616, (1960).
    5) Caspersson T., Zech L., Johansson C.: Differential banding of alkylating fluorochromes in human chromosomes. Exp. Cell. Res. 60(3), 315-319, (1970).
    6) Seabright M.: A rapid banding technique for human chromosomes. Lancet II:971-972 (1971).
    7) Yunis et al.: High resolution of human chromosomes. Science (1976) 191: 1268-1270.
    8) Webber L.M. and Garson O.M.: Fluoredeoxyuridine synchronization of bone marrow cultures. Cancer and Cytogenetics (1983) 8: 123-132.
    9) Genetic disorders and the fetus. Ed. A. Milunsky 3rd edition (1992).
    10) Chang H.-C., Jones O.W., Masui H.: Human amniotic fluid cells grown in a hormone-supplemented medium: Suitability for prenatal diagnosis. Proc. Natl. Acad. Sci. Vol. 79, 4795-4799, (August 1982).
    11) Brambati B., Simoni G.: Fetal diagnosis of trisomy 21 in the first trimester of pregnancy. Lancet (March 12, 1983).
    12) Simoni G., Brambati B., Danesino C., Terzoli G.L., Romitti L., Rossella F., Fraccaro M.: Diagnostic application of first trimester trophoblast sampling in 100 pregnancies. Hum. Genet. (1984) 64: 252-259.
    13) Fraccaro M., Simoni G., Brambati B.: First trimester fetal diagnosis. Springer-Verlag (1985)
    14) Gibas L.M., Grujic S., Barr M.A., Jackson L.G.: A simple technique for obtaining high quality chromosome preparations from chorionic villus samples using FdU synchronization. Prenat. Diagn. (1987) 7: 323-327.
    15) Lundsteen C., Lind A.M.: A test of climate room for preparation of chromosome slides. Clin. Genet. 23, 240-242, (1985).
    16) Spurbeck J.L., Zinsmeister A.R., Meyer J. and Jalal S.M.: Dynamics of chromosome spreading. American Journal of Medical Genetics 61:387-393, (1996).
    17) Claussen U., Michel S., Mühlig P., Westermann M., Grummt U.-W., Kromeyer-Hauschild K., and Liehr T.: Demistifying chromosome preparation and the implications for the concept of chromosome condensation during mitosis. Cytogenet. Genome Res. 98:136-146, (2002).
    18) Ami D., Di Segni M., Forcella M., Meraviglia V., Baccarin M., Doglia S.M., Terzoli G. Role of water in chromosome spreading and swelling induced by acetic acid treatment: a FTIR spectroscopy study. European Journal of Histochemistry: volume 58:2330, 2014.
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