Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Chromosomal in situ suppression hybridization of human gonosomes and autosomes and its use in clinical cytogenetics


DNA libraries from sorted human gonosomes were used selectively to stain the X and Y chromosomes in normal and aberrant cultured human cells by chromosomal in situ suppression (CISS-) hybridization. The entire X chromosome was stained in metaphase spreads. Interphase chromosome domains of both the active and inactive X were clearly delineated. CISS-hybridization of the Y chromosome resulted in the specific decoration of the euchromatic part (Ypter-q11), whereas the heterochromatic part (Yq12) remained unlabeled. The stained part of the Y chromosome formed a compact domain in interphase nuclei. This approach was applied to amniotic fluid cells containing a ring chromosome of unknown origin (47,XY; +r). The ring chromosome was not stained by library probes from the gonosomes, thereby suggesting its autosomal origin. The sensitivity of CISS-hybridization was demonstrated by the detection of small translocations and fragments in human lymphocyte metaphase spreads after irradiation with 60Co-gamma-rays. Lymphocyte cultures from two XX-males were investigated by CISS-hybridization with Y-library probes. In both cases, metaphase spreads demonstrated a translocation of Yp-material to the short arm of an X chromosome. The translocated Y-material could also be demonstrated directly in interphase nuclei. CISS-hybridization of autosomes 7 and 13 was used for prenatal diagnosis in a case with a known balanced translocation t(7;13) in the father. The same translocation was observed in amniotic fluid cells from the fetus. Specific staining of the chromosomes involved in such translocations will be particularly important, in the future, in cases that cannot be solved reliably by conventional chromosome banding alone.

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


  1. Bickmore WA, Cooke HJ (1987) Evolution of homologous sequences on the human X and Y chromosomes, outside of the meiotic pairing segment. Nucleic Acids Res 15:6261–6271

  2. Boué A, Gallano P (1984) A collaborative study of the segregation of inherited chromosome structural rearrangements in 1356 prenatal diagnoses. Prenat Diagn 4:45–67

  3. Burns J, Chan VTW, Jonasson JH, Fleming KA, Taylor S, McGee J O'D (1985) Sensitive system for visualizing biotinylated DNA probes hybridized in situ: rapid sex determination of intact cells. J Clin Pathol 38:4678–4686

  4. Cremer T, Cremer C, Baumann H, Luedtke EK, Sperling K, Teuber V, Zorn C (1982) Rabl's model of the interphase chromosome arrangement tested in Chinese hamster cells by premature chromosome condensation and laser-UV-microbeam experiments. Hum Genet 60:46–56

  5. Cremer T, Landegent JE, Brückner H, Scholl HP, Schardin M, Hager HD, Devilee P, Pearson PL, Ploeg M van der (1986) Detection of chromosome aberrations in the human interphase nucleus by visualization of specific target DNAs with radioactive and non-radioactive in situ hybridization techniques: diagnosis of trisomy 18 with probe L1. 84. Hum Genet 74:346–352

  6. Cremer T, Lichter P, Borden J, Ward DC, Manuelidis L (1988a) Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome-specific library probes. Hum Genet 80:235–246

  7. Cremer T, Tesin D, Hopman AHN, Manuelidis L (1988b) Rapid interphase and metaphase assessment of specific chromosomal changes in neuroectodermal tumor cells by in situ hybridization with chemically modified DNA probes. Exp Cell Res 176:199–220

  8. Cremer T, Popp S, Emmerich P, Lichter P, Cremer C (1990) Rapid metaphase detection of radiation-induced chromosome aberrations in human lymphocytes by chromosomal suppression in situ hybridization. Cytometry 11:110–118

  9. Crolla JA, Llerena JC Jr (1988) A mosaic 45,X/46,X,r(?) karyo-type investigated with X and Y centromere-specific probes using a non-autoradiographic in situ hybridization technique. Hum Genet 81:81–84

  10. Devilee P, Thierry RF, Kievits T, Kolluri R, Hopman AHN, Willard HF, Pearson PL, Cornelisse CJ (1988) Detection of chromosome aneuploidy in interphase nuclei from human primary breast tumors using chromosome-specific repetitive DNA probes. Cancer Res 48:5825–5830

  11. Emmerich P, Jauch A, Hofmann MC, Cremer T, Walt H (1989) Interphase cytogenetics in paraffin embedded sections from human testicular germ cell tumor xenografts and in corresponding cultured cells. Lab Invest 61:235–242

  12. Hopman AHN, Ramaekers FCS, Raap AK, Beck JLM, Devilee P, Ploeg M van der, Vooijs GP (1988) In situ hybridization as a tool to study numerical chromosome aberrations in solid bladder tumors. Histochemistry 89:307–316

  13. Hopman AHN, Ramaekers FCS, Vooijs GP (1989) Interphase cytogenetics on solid tumors. In: Polak JM, McGee J O'D (eds) In situ hybridization: principles and practice. Oxford University Press, Oxford, pp 165–186

  14. Landegent JE, Jansen in de Wal N, Ommen GJB van, De Baas F, Vijlder JJM, Duijn P van, Ploeg M van de (1985) Chromosomal localization of a unique gene by non-autoradiographic in situ hybridization. Nature 317:175–177

  15. Landegent JE, Jansen in de Wal N, Dirks RW, Baas F, Ploeg M van der (1987) Use of whole cosmid cloned genomic sequences for chromosomal localization by non-radioactive in situ hybridization. Hum Genet 77:366–370

  16. Langer PR, Waldrop AA, Ward DC (1981) Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci USA 78:6633–6637

  17. Lawrence JB, Villnave CA, Singer RH (1988) Sensitive, high-resolution chromatin and chromosome mapping in situ: presence and orientation of two closely integrated copies of EBV in a lymphoma line. Cell 52:51–61

  18. Lichter P, Cremer T, Borden J, Manuelidis L, Ward DC (1988a) Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries. Hum Genet 80:224–234

  19. Lichter P, Cremer T, Chang Tang CJ, Watkins PC, Manuelidis L, Ward DC (1988b) Rapid detection of chromosome 21 aberrations by in situ hybridization. Proc Natl Acad Sci USA 85:9664–9668

  20. Lichter P, Chang Tang CJ, Call K, Hermanson G, Evans GA, Housman D, Ward DC (1990) High resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. Science 247:63–69

  21. Maniatis T, Fritsch EF, Sambrook J (eds) (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

  22. Manuelidis L (1985) Individual interphase chromosome domains revealed by in situ hybridization. Hum Genet 71:288–293

  23. Nederlof PM, Robinson D, Abuknesha R, Wiegant J, Hopman AHN, Tanke HJ, Raap AK (1989) Three-color fluorescence in situ hybridization for the simultaneous detection of multiple nucleic acid sequences. Cytometry 10:20–27

  24. Pinkel D, Gray JW, Trask B, Engh G van den, Fuscoe J, Dekken H van (1986) Cytogenetic analysis by in situ hybridization with fluorescently labeled nucleic acid probes. Cold Spring Harb Symp Quant Biol 51:151–157

  25. Pinkel D, Landegent J, Collins C, Fusco J, Segraves R, Lucas J, Gray JW (1988) Fluorescence in situ hybridization with human chromosome specific libraries: detection of trisomy 21 and ranslocations of chromosome 4. Proc Natl Acad Sci USA 85:9138–9142

  26. Rappold G, Cremer T, Hager H-D, Davies KE, Müller CR, Yang T (1984) Sex chromosome positions in human interphase nuclei as studied by in situ hybridization with chromosome specific DNA-probes. Hum Genet 67:317–325

  27. Rooney DE, Czepulkowski BH (1986) Human cytogenetics: a practical approach. (Practical approach series) IRL Press, Oxford Washington, DC

  28. Sachs ES, Hemel JO van, Hollander JC den, Jahoda MGJ (1987) Marker chromosomes in a series of 10000 prenatal diagnoses. Cytogenetic and follow-up studies. Prenat Diagn 7:81–89

  29. Schardin M, Cremer T, Hager HD, Lang M (1985) Specific staining of human chromosomes in Chinese hamster × man hybrid cell lines demonstrates interphase chromosome territories. Hum Genet 71:281–287

  30. Schwarzacher HG, Wolf U (1970) Methoden in der medizinischen Zytogenetik. Springer, Berlin Heidelberg New York

  31. Steinbach P, Djalali M, Hansmann I, Kattner E, Meisel-Stosiek M, Probeck H-D, Schmidt A, Wolf M (1983) The genetic significance of accessory bisatellited marker chromosomes. Hum Genet 65:155–164

  32. Viegas-Péquignot E, Zhen Lin L. Dutrillaux B, Apiou F, Paulin D (1989) Assignment of human desmin gene to band 2q35 by nonradioactive in situ hybridization. Hum Genet 83:33–36

  33. Walt H, Emmerich P, Cremer T, Hofman M-C, Bannwart F (1989) Supernumerary chromosome 1 in interphase nuclei of atypical gern cells in paraffin-embedded human seminiferous tubules. Lab Invest 61:527–531

  34. Warburton D (1984) Outcome of cases of de novo structural rearrangements diagnosed at amniocentesis. Prenat Diagn 4:69–80

  35. Wienberg J, Jauch A, Stanyon R, Cremer T (1990) Molecular cytotaxonomy of primates by chromosomal in situ suppression hybridization. Genomics (in press)

Download references

Author information

Correspondence to A. Jauch.

Additional information

Dedicated to Professor Friedrich Vogel on the occasion of his 65th birthday

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Jauch, A., Daumer, C., Lichter, P. et al. Chromosomal in situ suppression hybridization of human gonosomes and autosomes and its use in clinical cytogenetics. Hum Genet 85, 145–150 (1990). https://doi.org/10.1007/BF00193186

Download citation


  • Prenatal Diagnosis
  • Interphase Nucleus
  • Chromosome Banding
  • Lymphocyte Culture
  • Metaphase Spread