Abstract
Fluorescence in situ hybridization (FISH) requires nucleic acid probes, including deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or nucleic acid analogs, labeled directly with fluorophores, or capable of indirect association with fluorophores. The nucleic acid provides the FISH assay with its specificity through complementary pairing of the probe nucleotides with nucleotides of the target nucleic acid. The appended fluorophores provide the ability to visually detect the homologous regions within the cellular structure using a fluorescence microscope. Photographic or electronic cameras can also be used to provide permanent images of the fluorescence staining patterns, and the latter can be used to provide quantitative measurements of the probe fluorescence.
Keywords
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.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Bueno D., Skinner J., Abud H., and Heath J. K. (1996) Double in situ hybridization on mouse embryos for detection of overlapping regions of gene expression. Trends Genet. 12, 385–387.
Diwu Z., Klaubert D. H., and Haugland R. P. (1999) Spectral properties and biological applications of ELF enzyme substrates that generate bright fluorescent precipitates at the enzymatic activity sites. Proc. SPIE-Intl. Soc. Opt. Eng. 3602, 265.
Jowett T. and Yan Y. L. (1996) Double fluorescent in situ hybridization to zebrafish embryos. Trends Genet. 12, 387–389.
Bobrow M. N., Harris T. D., Shaughnessy K. J., and Litt G. J. (1989) Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays. J. Immunol. Methods 125, 279–285.
Raap A. K., Van De Corput M. P., Vervenne R. A., Van Gijlswijk R. P., Tanke H. J., and Wiegant J. (1995) Ultra-sensitive FISH using peroxidase-mediated deposition of biotin-or fluorochrome tyramides. Hum. Mol. Genet. 4, 529–534.
Speel E. J., Ramaekers F. C., and Hopman A. H. (1997) Sensitive multicolor fluorescence in situ hybridization using catalyzed reporter deposition (CARD) amplification. J. Histochem. Cytochem. 45, 1439–1446.
Bauman J. G., and Van Duijn P. (1981) Hybrido-cytochemical localization of specific DNA sequences by fluorescence microscopy. Histochem. J. 13, 723–733.
Cremer T., Landegent J., Bruckner A., et al. (1986) Detection of chromosome aberrations in the human interphase nucleus by visualization of specific target DNAs with radioactive and nonradioactive in situ hybridization techniques: Diagnosis of trisomy 18 with probe L1.84. Hum. Genet. 74, 346–352.
Cremer T., Tesin D., Hopman A. H., and Manuelidis L. (1988) 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.
Landegent J. E., Jasen in De Wal N., Baan R. A., Hoeijmakers J. H., and Van Der Ploeg M. (1984) 2-Acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences. Exp. Cell Res. 153, 61–72.
Nederlof P. M., Robinson D., Abuknesha R., et al. (1989) Three-color fluorescence in situ hybridization for the simultaneous detection of multiple nucleic acid sequences. Cytometry 10, 20–27.
Nederlof P. M., Van Der Flier S., Wiegant J., et al. (1990) Multiple fluorescence in situ hybridization. Cytometry 11, 126–131.
Dale R. M. and Ward D. C. (1975) Mercurated polynucleotides: New probes for hybridization and selective polymer fractionation. Biochemistry 14, 2458–2469.
Hopman A. H., Wiegant J., Tesser G. I., and Van Duijn P. (1986) A nonradioactive in situ hybridization method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands. Nucl. Acids Res. 14, 6471–6488.
Hopman A. H., Wiegant J., and Van Duijn P. (1987) Mercurated nucleic acid probes, a new principle for nonradioactive in situ hybridization. Exp. Cell Res. 169, 357–368.
Van Belkum A., Linkels E., Jelsma T., Van Den Berg F. M., and Quint W. (1994) Nonisotopic labeling of DNA by newly developed hapten-containing platinum compounds. Biotechniques 16, 148–153.
Draper D. E. (1984) Attachment of reporter groups to specific, selected cytidine residues in RNA using a bisulfite-catalyzed transamination reaction. Nucl. Acids Res. 12, 989–1002.
Reisfeld A., Rothenberg J. M., Bayer E. A., and Wilchek M. (1987) Nonradioactive hybridization probes prepared by the reaction of biotin hydrazide with DNA. Biochem. Biophys. Res. Commun. 142, 519–526.
Keller G. H., Huang D. P., and Manak M. M. (1989) Labeling of DNA probes with a photoactivatable hapten. Anal. Biochem. 177, 392–395.
Keller G. H., Cumming C. U., Huang D. P., Manak M. M., and Ting R. (1988) A chemical method for introducing haptens onto DNA probes. Anal. Biochem. 170, 441–450.
Chu B. C., Wahl G. M., and Orgel L. E. (1983) Derivatization of unprotected polynucleotides. Nucl. Acids Res. 11, 6513–6529.
Morrison L. E., Halder T. C., and Stols L. M. (1989) Solution-phase detection of polynucleotides using interacting fluorescent labels and competitive hybridization. Anal. Biochem. 183, 231–244.
Langer P. R., Waldrop A. A., and Ward D. C. (1981) Enzymatic synthesis of biotinlabeled polynucleotides: novel nucleic acid affinity probes. Proc. Natl. Acad. Sci. USA 78, 6633–6637.
Langer-Safer P. R., Levine M., and Ward D. C. (1982) Immunological method for mapping genes on Drosophila polytene chromosomes. Proc. Natl. Acad. Sci. USA 79, 4381–4385.
Manuelidis L., Langer-Safer P. R., and Ward D. C. (1982) High-resolution mapping of satellite DNA using biotin-labeled DNA probes. J. Cell. Biol. 95, 619–625.
Ried T., Landes G., Dackowski W., Klinger K., and Ward D. C. (1992) Multicolor fluorescence in situ hybridization for the simultaneous detection of probe sets for chromosomes 13, 18, 21, X and Y in uncultured amniotic fluid cells. Hum. Mol. Genet. 1, 307–313.
Arnoldus E. P., Wiegant J., Noordermeer I. A., et al. (1990) Detection of the Philadelphia chromosome in interphase nuclei. Cytogenet. Cell. Genet. 54, 108–111.
Wiegant J., Ried T., Nederlof P. M., Van Der Ploeg M., Tanke H. J., and Raap A. K. (1991) In situ hybridization with fluoresceinated DNA. Nucl. Acids Res. 19, 3237–3241.
Ried T., Baldini A., Rand T. C., and Ward D. C. (1992) Simultaneous visualization of seven different DNA probes by in situ hybridization using combinatorial fluorescence and digital imaging microscopy. Proc. Natl. Acad. Sci. USA 89, 1388–1392.
Telenius H., Pelmear A. H., Tunnacliffe A., et al. (1992) Cytogenetic analysis by chromosome painting using DOP-PCR amplified flow-sorted chromosomes. Genes Chromosomes Cancer 4, 257–263.
Morrison L. E. and Legator M. S. (1999) Multi-color fluorescence in situ hybridizations techniques, in An Introduction to Fluorescence in situ Hybridization: Principles and Clinical Applications (eiAndreeff M. and Pinkel D., eds.), Wiley-Liss, New York, pp. 77–118.
Sambrook J., Fritsch E. F., and Maniatis T. (1989) Molecular Cloning, A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.
Collins C., Kuo W. L., Segraves R., Fuscoe J., Pinkel D., and Gray J. W. (1991) Construction and characterization of plasmid libraries enriched in sequences from single human chromosomes. Genomics 11, 997–1006.
Deaven L. L., Van Dilla M. A., Bartholdi M. F., et al. (1986) Construction of human chromosome-specific DNA libraries from flow-sorted chromosomes. Cold Spring Harb. Symp. Quant. Biol. 51, 159–167.
Carter N. P., Ferguson-Smith M. A., Perryman M. T., et al. (1992) Reverse chromosome painting: A method for the rapid analysis of aberrant chromosomes in clinical cytogenetics. J. Med. Genet. 29, 299–307.
Telenius H., De Vos D., Blennow E., Willat L. R., Ponder B. A., and Carter N. P. (1993) Chromatid contamination can impair the purity of flow sorted metaphase chromosomes. Cytometry 14, 97–101.
Guan X. Y., Meltzer P. S., and Trent J. M. (1994) Rapid generation of whole chromosome painting probes (WCPs) by chromosome microdissection. Genomics 22, 101–107.
Telenius H., Carter N. P., Bebb C. E., Nordenskjold M., Ponder B. A., and Tunnacliffe A. (1992) Degenerate oligonucleotide-primed PCR: General amplification of target DNA by a single degenerate primer. Genomics 13, 718–725.
Cherif D., Bernard O., and Berger R. (1989) Detection of single-copy genes by nonisotopic in situ hybridization on human chromosomes. Hum. Genet. 81, 358–362.
Kelly R. B., Cozzarelli N. R., Deutscher M. P., Lehman I. R., and Kornberg A. (1970) Enzymatic synthesis of deoxyribonucleic acid. XXXII. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break. J. Biol. Chem. 245, 39–45.
Feinberg A. P. and Vogelstein B. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13.
Feinberg A. P. and Vogelstein B. (1984) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Addendum. Anal. Biochem. 137, 266–267.
Speicher M. R., Gwyn Ballard S., and Ward D. C. (1996) Karyotyping human chromosomes by combinatorial multi-fluor FISH. Nat. Genet. 12, 368–375.
Schrock E., Du Manoir S., Veldman T., et al. (1996) Multicolor spectral karyotyping of human chromosomes. Science 273, 494–497.
Morrison L., Legator M., Ramakrishnan R., Zimmerman D., Volling-Kwasneski M., and Koeppen P. (1998) FISH karyotyping with combinations of whole chromosome probes directly labeled with five visible fluorescent labels. Cytometry Supplement 9, 150.
Morrison L. E. (1995) Detection of energy transfer and fluorescence quenching, in Nonisotopic probing, blotting, and sequencing (Kricka L. J., ed.), Academic Press, San Diego, pp. 429–471.
Van Belkum A., Linkels E., Jelsma T., Houthoff H. J., Van Den Berg F., and Quint W. (1993) Application of a new, universal DNA labeling system in the PCR mediated diagnoses of Chlamydia trachomatis and human papillomavirus type 16 infection in cervical smears. J. Virol. Methods 45, 189–200.
Garvey J. S., Cremer N. E., and Sussdorf D. H. (1977) Methods in Immunology: A Laboratory Text for Instruction and Research. 3rd ed. W. A. Benjamin, Inc., Reading, PA.
Morrison L. E. (1988) Time-resolved detection of energy transfer: Theory and application to immunoassays. Anal. Biochem. 174, 101–120.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc.
About this protocol
Cite this protocol
Morrison, L.E., Ramakrishnan, R., Ruffalo, T.M., Wilber, K.A. (2002). Labeling Fluorescence In Situ Hybridization Probes for Genomic Targets. In: Fan, YS. (eds) Molecular Cytogenetics. Methods in Molecular Biology™, vol 204. Humana Press. https://doi.org/10.1385/1-59259-300-3:21
Download citation
DOI: https://doi.org/10.1385/1-59259-300-3:21
Publisher Name: Humana Press
Print ISBN: 978-1-58829-006-9
Online ISBN: 978-1-59259-300-2
eBook Packages: Springer Protocols
