• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. May 1986; 83(9): 2934–2938.
PMCID: PMC323421

Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization.

Abstract

This report describes the use of fluorescence in situ hybridization for chromosome classification and detection of chromosome aberrations. Biotin-labeled DNA was hybridized to target chromosomes and subsequently rendered fluorescent by successive treatments with fluorescein-labeled avidin and biotinylated anti-avidin antibody. Human chromosomes in human-hamster hybrid cell lines were intensely and uniformly stained in metaphase spreads and interphase nuclei when human genomic DNA was used as a probe. Interspecies translocations were detected easily at metaphase. The human-specific fluorescence intensity from cell nuclei and chromosomes was proportional to the amount of target human DNA. Human Y chromosomes were fluorescently stained in metaphase and interphase nuclei by using a 0.8-kilobase DNA probe specific for the Y chromosome. Cells from males were 40 times brighter than those from females. Both Y chromosomal domains were visible in most interphase nuclei of XYY amniocytes. Human 28S ribosomal RNA genes on metaphase chromosomes were distinctly stained by using a 1.5-kilobase DNA probe.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.4M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Caspersson T, Farber S, Foley GE, Kudynowski J, Modest EJ, Simonsson E, Wagh U, Zech L. Chemical differentiation along metaphase chromosomes. Exp Cell Res. 1968 Jan;49(1):219–222. [PubMed]
  • Nilsson B. A bibliography of literature concerning chromosome identification--with special reference to fluorescence Giemsa staining techniques. Hereditas. 1973;73(2):259–270. [PubMed]
  • Burgerhout W. Identification of interspecific translocation chromosomes in human-Chinese hamster hybrid cells. Humangenetik. 1975 Sep 23;29(3):229–231. [PubMed]
  • Langer PR, Waldrop AA, Ward DC. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633–6637. [PMC free article] [PubMed]
  • Landegent JE, Jasen in de Wal N, Baan RA, Hoeijmakers JH, Van der Ploeg M. 2-Acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences. Exp Cell Res. 1984 Jul;153(1):61–72. [PubMed]
  • Bauman JG, Wiegant J, Borst P, van Duijn P. A new method for fluorescence microscopical localization of specific DNA sequences by in situ hybridization of fluorochromelabelled RNA. Exp Cell Res. 1980 Aug;128(2):485–490. [PubMed]
  • Zorn C, Cremer C, Cremer T, Zimmer J. Unscheduled DNA synthesis after partial UV irradiation of the cell nucleus. Distribution in interphase and metaphase. Exp Cell Res. 1979 Nov;124(1):111–119. [PubMed]
  • Devine EA, Nolin SL, Houck GE, Jr, Jenkins EC, Brown WT. Chromosomal localization of several families of repetitive sequences by in situ hybridization. Am J Hum Genet. 1985 Jan;37(1):114–123. [PMC free article] [PubMed]
  • Lau YF, Schonberg S. A male-specific DNA probe detects heterochromatin sequences in a familial Yq- chromosome. Am J Hum Genet. 1984 Nov;36(6):1394–1396. [PMC free article] [PubMed]
  • Graham GJ, Hall TJ, Cummings MR. Isolation of repetitive DNA sequences from human chromosome 21. Am J Hum Genet. 1984 Jan;36(1):25–35. [PMC free article] [PubMed]
  • Yang TP, Hansen SK, Oishi KK, Ryder OA, Hamkalo BA. Characterization of a cloned repetitive DNA sequence concentrated on the human X chromosome. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6593–6597. [PMC free article] [PubMed]
  • Willard HF. Chromosome-specific organization of human alpha satellite DNA. Am J Hum Genet. 1985 May;37(3):524–532. [PMC free article] [PubMed]
  • Jabs EW, Wolf SF, Migeon BR. Characterization of a cloned DNA sequence that is present at centromeres of all human autosomes and the X chromosome and shows polymorphic variation. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4884–4888. [PMC free article] [PubMed]
  • Gusella JF, Keys C, VarsanyiBreiner A, Kao FT, Jones C, Puck TT, Housman D. Isolation and localization of DNA segments from specific human chromosomes. Proc Natl Acad Sci U S A. 1980 May;77(5):2829–2833. [PMC free article] [PubMed]
  • Gusella JF, Keys C, VarsanyiBreiner A, Kao FT, Jones C, Puck TT, Housman D. Isolation and localization of DNA segments from specific human chromosomes. Proc Natl Acad Sci U S A. 1980 May;77(5):2829–2833. [PMC free article] [PubMed]
  • Thompson LH, Mooney CL, Burkhart-Schultz K, Carrano AV, Siciliano MJ. Correction of a nucleotide-excision-repair mutation by human chromosome 19 in hamster-human hybrid cells. Somat Cell Mol Genet. 1985 Jan;11(1):87–92. [PubMed]
  • Gray JW, Langlois RG. Chromosome classification and purification using flow cytometry and sorting. Annu Rev Biophys Biophys Chem. 1986;15:195–235. [PubMed]
  • Harper ME, Ullrich A, Saunders GF. Localization of the human insulin gene to the distal end of the short arm of chromosome 11. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4458–4460. [PMC free article] [PubMed]
  • Erickson JM, Rushford CL, Dorney DJ, Wilson GN, Schmickel RD. Structure and variation of human ribosomal DNA: molecular analysis of cloned fragments. Gene. 1981 Dec;16(1-3):1–9. [PubMed]
  • Van Prooijen-Knegt AC, Van Hoek JF, Bauman JG, Van Duijn P, Wool IG, Van der Ploeg M. In situ hybridization of DNA sequences in human metaphase chromosomes visualized by an indirect fluorescent immunocytochemical procedure. Exp Cell Res. 1982 Oct;141(2):397–407. [PubMed]
  • Duhamel RC, Johnson DA. Use of nonfat dry milk to block nonspecific nuclear and membrane staining by avidin conjugates. J Histochem Cytochem. 1985 Jul;33(7):711–714. [PubMed]
  • Johnson GD, Nogueira Araujo GM. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981;43(3):349–350. [PubMed]
  • Durnam DM, Gelinas RE, Myerson D. Detection of species specific chromosomes in somatic cell hybrids. Somat Cell Mol Genet. 1985 Nov;11(6):571–577. [PubMed]
  • Schardin M, Cremer T, Hager HD, Lang M. Specific staining of human chromosomes in Chinese hamster x man hybrid cell lines demonstrates interphase chromosome territories. Hum Genet. 1985;71(4):281–287. [PubMed]
  • Mendelsohn ML, Mayall BH, Bogart E, Moore DH, 2nd, Perry BH. DNA content and DNA-based centromeric index of the 24 human chromosomes. Science. 1973 Mar 16;179(4078):1126–1129. [PubMed]
  • Schmickel RD. Quantitation of human ribosomal DNA: hybridization of human DNA with ribosomal RNA for quantitation and fractionation. Pediatr Res. 1973 Jan;7(1):5–12. [PubMed]
  • Landegent JE, Jansen in de Wal N, van Ommen GJ, Baas F, de Vijlder JJ, van Duijn P, Van der Ploeg M. Chromosomal localization of a unique gene by non-autoradiographic in situ hybridization. Nature. 1985 Sep 12;317(6033):175–177. [PubMed]
  • Manuelidis L, Ward DC. Chromosomal and nuclear distribution of the HindIII 1.9-kb human DNA repeat segment. Chromosoma. 1984;91(1):28–38. [PubMed]
  • Albertson DG. Mapping muscle protein genes by in situ hybridization using biotin-labeled probes. EMBO J. 1985 Oct;4(10):2493–2498. [PMC free article] [PubMed]
  • Lau YF, Ying KL, Donnell GN. Identification of a case of Y:18 translocation using a Y-specific repetitive DNA probe. Hum Genet. 1985;69(2):102–105. [PubMed]
  • Müller CR, Davies KE, Cremer C, Rappold G, Gray JW, Ropers HH. Cloning of genomic sequences from the human Y chromosome after purification by dual beam flow sorting. Hum Genet. 1983;64(2):110–115. [PubMed]
  • Krumlauf R, Jeanpierre M, Young BD. Construction and characterization of genomic libraries from specific human chromosomes. Proc Natl Acad Sci U S A. 1982 May;79(9):2971–2975. [PMC free article] [PubMed]
  • Davies KE, Young BD, Elles RG, Hill ME, Williamson R. Cloning of a representative genomic library of the human X chromosome after sorting by flow cytometry. Nature. 1981 Oct 1;293(5831):374–376. [PubMed]
  • Disteche CM, Kunkel LM, Lojewski A, Orkin SH, Eisenhard M, Sahar E, Travis B, Latt SA. Isolation of mouse x-chromosome specific DNA from an x-enriched lambda phage library derived from flow sorted chromosomes. Cytometry. 1982 Mar;2(5):282–286. [PubMed]
  • Lalande M, Dryja TP, Schreck RR, Shipley J, Flint A, Latt SA. Isolation of human chromosome 13-specific DNA sequences cloned from flow sorted chromosomes and potentially linked to the retinoblastoma locus. Cancer Genet Cytogenet. 1984 Dec;13(4):283–295. [PubMed]
  • Rappold GA, Cremer T, Hager HD, Davies KE, Müller CR, Yang T. Sex chromosome positions in human interphase nuclei as studied by in situ hybridization with chromosome specific DNA probes. Hum Genet. 1984;67(3):317–325. [PubMed]
  • Manuelidis L. Different central nervous system cell types display distinct and nonrandom arrangements of satellite DNA sequences. Proc Natl Acad Sci U S A. 1984 May;81(10):3123–3127. [PMC free article] [PubMed]
  • Mathog D, Hochstrasser M, Gruenbaum Y, Saumweber H, Sedat J. Characteristic folding pattern of polytene chromosomes in Drosophila salivary gland nuclei. Nature. 308(5958):414–421. [PubMed]
  • Trask B, van den Engh G, Landegent J, in de Wal NJ, van der Ploeg M. Detection of DNA sequences in nuclei in suspension by in situ hybridization and dual beam flow cytometry. Science. 1985 Dec 20;230(4732):1401–1403. [PubMed]
  • Cooke HJ, Schmidtke J, Gosden JR. Characterisation of a human Y chromosome repeated sequence and related sequences in higher primates. Chromosoma. 1982;87(5):491–502. [PubMed]
  • Burns J, Chan VT, Jonasson JA, Fleming KA, Taylor S, McGee JO. Sensitive system for visualising biotinylated DNA probes hybridised in situ: rapid sex determination of intact cells. J Clin Pathol. 1985 Oct;38(10):1085–1092. [PMC free article] [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Cited in Books
    Cited in Books
    PubMed Central articles cited in books
  • Compound
    Compound
    PubChem Compound links
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...