Color-Changing Karyotyping (CCK), an M-FISH/SKY Alternative

  • Octavian Henegariu
Part of the Springer Protocols Handbooks book series (SPH)

Multicolor karyotyping of human chromosomes is a valuable tool for defining marker chromosomes as well as small, cryptic or complex chromosome translocations. Two procedures, multiplex FISH (M-FISH) and spectral karyotyping (SKY) use combinatorial labeling schemes involving 5–6 different fluorophores to label DNA painting probes specific for all human (or mouse) chromosomes. Although the results obtained are excellent, both procedures require expensive equipment for detection and specialized software for analysis. In many laboratories, research projects often only occasionally require multicolor karyotyping, and so expensive investment in specialized accessory hardware cannot be justified. To minimize costs and yet enable color karyotyping using only the most basic equipment, we have developed an alternative technique called color-changing karyotyping (CCK), which uses DNA probes labeled with only three fluorescent dyes. Image capture can be accomplished using a conventional digital camera, and image processing can be performed with generic imaging software. An optimized procedure for cytogenetic slide preparation and efficient chromosome hybridization is also detailed.


Painting Probe Metal Block Fish Signal Automatic Pipette Chromosome Painting Probe 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Bains W (1991) Hybridization methods for DNA sequencing. Genomics 11:294–301CrossRefPubMedGoogle Scholar
  2. du Manoir S, Schrock E, Bentz M, Speicher MR, Joos S, Ried T, Lichter P, Cremer T (1995) Quantitative analysis of comparative genomic hybridization. Cytometry 19:27–41CrossRefPubMedGoogle Scholar
  3. Haas M, Aburatani H, Stanton VP, Jr., Bhatt M, Housman D, Ward DC (1993) Isolation and FISH mapping of 80 cosmid clones on the short arm of human chromosome 3. Genomics 16: 90–96CrossRefPubMedGoogle Scholar
  4. Henegariu O, Heerema NA, Bray-Ward P, Ward DC (1999) Colour-changing karyotyping: an alternative to M-FISH/SKY. Nat Genet 23:263–264CrossRefPubMedGoogle Scholar
  5. Horsley SW, Mackay A, Iravani M, Fenwick K, Valgeirsson H, Dexter T, Ashworth A, Kearney L (2006) Array CGH of fusion gene-positive leukemia-derived cell lines reveals cryptic regions of genomic gain and loss. Genes Chromosomes Cancer 45:554–564CrossRefPubMedGoogle Scholar
  6. Krause DS, Theise ND, Collector MI, Henegariu O, Hwang S, Gardner R, Neutzel S, Sharkis SJ (2001) Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell. Cell 105:369–377CrossRefPubMedGoogle Scholar
  7. Liehr T, Starke H, Weise A, Lehrer H, Claussen U (2004) Multicolor FISH probe sets and their applications. Histol Histopathol 19:229–237PubMedGoogle Scholar
  8. Ozturk M, Bolkent S, Yilmazer S, Kaner G, Unal H (1998) Detection of c-erbB-2 mRNAs using dig-labelled oligonucleotide probe with in situ hybridisation in human breast carcinoma: comparison with immunohistochemical results. Anal Cell Pathol 16:201–209PubMedGoogle Scholar
  9. Schrück E, du Manoir S, Veldman T, Schoell B, Wienberg J, Ferguson-Smith MA, Ning Y, Ledbetter DH, Bar-Am I, Soenksen D, Garini Y, Ried T (1996) Multicolor spectral karyotyping of human chromosomes [see comments]. Science 273:494–497CrossRefGoogle Scholar
  10. Speicher MR, Gwyn Ballard S, Ward DC (1996) Karyotyping human chromosomes by combinatorial multi-fluor FISH. Nat Genet 12:368–375CrossRefPubMedGoogle Scholar
  11. Theise ND, Henegariu O, Grove J, Jagirdar J, Kao PN, Crawford JM, Badve S, Saxena R, Krause DS (2002) Radiation pneumonitis in mice: a severe injury model for pneumocyte engraftment from bone marrow. Exp Hematol 30:1333–1338CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2009

Authors and Affiliations

  • Octavian Henegariu
    • 1
  1. 1.Department of ImmunobiologyYale UniversityNew HavenUSA

Personalised recommendations