Multicolor fluorescence in situ hybridization (mFISH) assays are now indispensable for precisely describing complex chromosomal rearrangements and marker chromosomes. The routine application of such techniques to human chromosomes began in 1996 with the simultaneous use of all 24 human whole-chromosome painting (wcp) probes in multiplex FISH (M-FISH) and spectral karyotyping (SKY). This chapter reviews the available mFISH approaches that use wcp probes, and describes basic protocols for M-FISH and SKY.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Aurich-Costa J, Vannier A, Gregoire E, Nowak F, Cherif D (2001) IPM-FISH, a new M-FISH approach using IRS-PCR painting probes: Application to the analysis of seven human prostate cell lines. Genes Chromosomes Cancer 30:143–160
Azofeifa J, Fauth C, Kraus J, Maierhofer C, Langer S, Bolzer A, Reichman J, Schuffenhauer S, Speicher MR (2000) An optimized probe set for the detection of small interchromosomal aberrations by use of 24-color FISH. Am J Hum Genet 66:1684–1688
Buwe A, Steinlein C, Koehler MR, Bar-Am I, Katzin N, Schmid M (2003) Multicolor spectral karyotyping of rat chromosomes. Cytogenet Genome Res 103:163–168
Chang SS, Mark HF (1997) Emerging molecular cytogenetic technologies. Cytobios 90:7–22
Claussen U, Michel S, Mühlig P, Westermann M, Grummt UW, Kromeyer-Hauschild K, Liehr T (2002) Demystifying chromosome preparation and the implications for the concept of chromosome condensation during mitosis. Cytogenet Genome Res 98:136–146
Griffin DK, Haberman F, Masabanda J, O'Brien P, Bagga M, Sazanov A, Smith J, Burt DW, Ferguson-Smith M, Wienberg J (1999) Micro- and macrochromosome paints generated by flow cytometry and microdissection: Tools for mapping the chicken genome. Cytogenet Cell Genet 87:278–281
Karst C, Trifonov V, Romanenko SA, Claussen U, Mrasek K, Michel S, Avner P, Liehr T (2006) Molecular cytogenetic characterization of the mouse cell line WMP2 by spectral karyotyping and multicolor banding applying murine probes. Int J Mol Med 17:209–213
Liehr T (2008) Multicolor FISH homepage. http://www.med.uni-jena.de/fish/mFISH/mFISHlit.htm
Liehr T, Claussen U (2002) Multicolor-FISH approaches for the characterization of human chromosomes in clinical genetics and tumor cytogenetics. Curr Genom 3:213–235
Liehr T, Starke H, Weise A, Lehrer H, Claussen U (2004) Multicolor FISH probe sets and their applications. Histol Histopathol 19:229–237
Liyanage M, Coleman A, du Manoir S, Veldman T, McCormack S, Dickson RB, Barlow C, Wynshaw-Boris A, Janz S, Wienberg J, Ferguson-Smith MA, Schröck E, Ried T (1996) Multicolour spectral karyotyping of mouse chromosomes. Nat Genet 14:312–315
Milne SB, Hoather T, O'Brien PCM, Yang F, Ferguson-Smith MA, Dobson J, Sargan D (2004) Karyotype of canine soft tissue sarcomas: A multi-colour, multi-species approach to canine chromosome painting. Chromosome Res 12:825–835
Mrasek K, Heller A, Rubtsov N, Trifonov V, Starke H, Rocchi M, Claussen U, Liehr T (2001) Reconstruction of the female Gorilla gorilla karyotype using 25-color FISH and multicolor banding (MCB). Cytogenet Cell Genet 93:242–248
Nederlof PM, Robinson D, Abuknesha R, Wiegant J, Hopman AH, Tanke HJ, Raap AK (1989) Three-color fluorescence in situ hybridization for the simultaneous detection of multiple nucleic acid sequences. Cytometry 10:20–27
Rutten KB, Pietsch C, Olek K, Neusser M, Beukeboom LW, Gadau J (2004) Chromosomal anchoring of linkage groups and identification of wing size QTL using markers and FISH probes derived from microdissected chromosomes in Nasonia (Pteromalidae: Hymenoptera). Cytogenet Genome Res 105:126–133
Schröck 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. Science 273:494–497
Senger G, Chudoba I, Plesch A (1998) Multicolor-FISH—the identification of chromosome aberrations by 24 colors. BIOforum 9:499–503
Speicher MR, Gwyn Ballard S, Ward DC (1996) Karyotyping human chromosomes by combinatorial multi-fluor FISH. Nat Genet 12:368–375
Szuhai K, Sandhaus E, Kolkman-Uljee SM, Lemaitre M, Truffert JC, Dirks RW, Tanke HJ, Fleuren GJ, Schuuring E, Raap AK (2001) A novel strategy for human papillomavirus detection and genotyping with SybrGreen and molecular beacon polymerase chain reaction. Am J Pathol 159:1651–1660
Tanke HJ, De Haas RR, Sagner G, Ganser M, van Gijlswijk RP (1998) Use of platinum copropor-phyrin and delayed luminescence imaging to extend the number of targets FISH karyotyping. Cytometry 33:453–459
Tanke HJ, Wiegant J, van Gijlswijk RP, Bezrookove V, Pattenier H, Heetebrij RJ, Talman EG, Raap AK, Vrolijk J (1999) New strategy for multi-colour fluorescence in situ hybridisation: COBRA: COmbined Binary RAtio labelling. Eur J Hum Genet 7:2–11
Tosi S, Giudici G, Rambaldi A, Scherer SW, Bray-Ward P, Dirscherl L, Biondi A, Kearney L (1999) Characterization of the human myeloid leukemia-derived cell line GF-D8 by multiplex fluorescence in situ hybridization, subtelomeric probes, and comparative genomic hybridization. Genes Chromosomes Cancer 24:213–221
Yurov YB, Soloviev IV, Vorsanova SG, Marcais B, Roizes G, Lewis R (1996) High resolution multicolor fluorescence in situ hybridization using cyanine and fluorescein dyes: Rapid chromosome identification by directly fluorescently labeled alphoid DNA probes. Hum Genet 97:390–398
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Springer-Verlag Berlin Heidelberg
About this protocol
Cite this protocol
Liehr, T. et al. (2009). Multiplex FISH and Spectral Karyotyping. In: Liehr, T. (eds) Fluorescence In Situ Hybridization (FISH) — Application Guide. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-70581-9_17
Download citation
DOI: https://doi.org/10.1007/978-3-540-70581-9_17
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-540-70580-2
Online ISBN: 978-3-540-70581-9
eBook Packages: Springer Protocols