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Human Genetics

, Volume 97, Issue 3, pp 390–398 | Cite as

High resolution multicolor fluorescence in situ hybridization using cyanine and fluorescein dyes: Rapid chromosome identification by directly fluorescently labeled alphoid DNA probes

  • Yuri B. Yurov
  • Ilia V. Soloviev
  • Svetlana G. Vorsanova
  • Bertrand Marcais
  • Gerard Roize
  • Robert Lewis
Original Investigation

Abstract

We tested DNA probes directly labeled by fluorescently labeled nucleotides (Cy3-dCTP, Cy5-dCTP, FluorX-dCTP) for high resolution uni- and multicolor detection of human chromosomes and analysis of centromeric DNA organization by in situ hybridization. Alpha-satellite DNA probes specific to chromosomes 1, 2, 3, 4 + 9, 5 + 19, 6, 7, 8, 10, 11, 13 + 21, 14 + 22, 15, 16, 17, 18, 20, 22, X and Y were suitable for the accurate identification of human chromosomes in metaphase and interphase cells. Cy3-labeled probes had several advantages: (1) a high level of fluorescence (5–10 times more compared with fluorescein-labeled probes); (2) a low level of fluorescence in solution, allowing the detection of target chromosomes in situ during hybridization without the washing of slides; and (3) high resistance to photobleaching during prolonged (1-2 h) exposure to strong light, thus allowing the use of a high energy mercury lamp or a long integration time during image acquisition in digital imaging microscopy for the determination of weak signals. For di- and multicolor fluorescence in situ hybridization (FISH), we successfully used different combinations of directly fluorophorated probes with preservation of images by conventional microscopy or by digital imaging microscopy. FluorX and Cy3 dyes allowed the use of cosmid probes for mapping in a one-step hybridization experiment. Cyanine-labeled fluorophorated DNA probes offer additional possibilities for rapid chromosome detection during a simple 15-min FISH procedure, and can be recommended for basic research and clinical studies, utilizing FISH.

Keywords

Cyanine Target Chromosome Digital Imaging Microscopy Multicolor Fluorescence Cosmid 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.

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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Yuri B. Yurov
    • 1
    • 2
    • 3
  • Ilia V. Soloviev
    • 1
  • Svetlana G. Vorsanova
    • 2
    • 3
  • Bertrand Marcais
    • 3
  • Gerard Roize
    • 3
  • Robert Lewis
    • 4
  1. 1.National Research Centre of Mental HealthRussian Academy of Medical Sciences, MoscowMoscowRussia
  2. 2.Insitute of Pediatrics and Children SurgeryRussian Ministry of HealthMoscowRussia
  3. 3.Institut de BiologieINSERM U249-CNRSMontpellierFrance
  4. 4.Biological Detection Systems, Inc.PittsburghUSA

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