Three-Dimensional Fluorescence In Situ Hybridization in Mouse Embryos Using Repetitive Probe Sequences

  • Walid E. MaaloufEmail author
  • Tiphaine Aguirre-Lavin
  • Laetitia Herzog
  • Isabelle Bataillon
  • Pascale Debey
  • Nathalie Beaujean
Part of the Methods in Molecular Biology book series (MIMB, volume 659)


A common problem in research laboratories that study the mammalian embryo is the limited supply of live material. For this reason, new methods are constantly being developed and existing methods for in vitro models using cells in culture are being adapted to represent embryogenesis. Three-dimensional fluorescence in situ hybridization (3D-FISH) is an important tool to study where genomic sequences are positioned within nuclei without interfering with this 3D organization. When used in the embryo, this technique provides vital information about the distribution of specific sequences in relation to embryonic nuclear substructures such as nucleolar precursor bodies and chromocenters. In this chapter, we will present a detailed description of FISH in order to perform 3D-FISH in the early preimplantation murine embryos.

Key words

3-Dimensional FISH In situ Embryo Mouse Development 



We thank Pierre Adenot for technical assistance on the confocal microscope platform MIMA2 (Microscopie et Imagerie des Microorganismes, Animaux et Elements) and UEAR for animal care. We are also grateful to Eve Devinoy (INRA) and Claire Francastel (Institut Cochin) for their help. This work was supported by INRA grant “Crédits incitatifs PHASE”.


  1. 1.
    Halling, K. C., Kipp, B. R. (2007) Fluorescence in situ hybridization in diagnostic cytology. Hum Pathol 38, 1137–1144.PubMedCrossRefGoogle Scholar
  2. 2.
    Sreekantaiah, C. (2007) FISH panels for hematologic malignancies. Cytogenet Genome Res 118, 284–296.PubMedCrossRefGoogle Scholar
  3. 3.
    Foster, H. A., Bridger, J. M. (2005) The genome and the nucleus: a marriage made by evolution. Genome organisation and nuclear architecture. Chromosoma 114, 212–229.PubMedCrossRefGoogle Scholar
  4. 4.
    Walter, J., Joffe, B., Bolzer, A., Albiez, H., Benedetti, P. A., Muller, S., Speicher, M. R., Cremer, T., Cremer, M., Solovei, I. 2006 Towards many colors in FISH on 3D-preserved interphase nuclei. Cytogenet Genome Res 114, 367–378.PubMedCrossRefGoogle Scholar
  5. 5.
    Kupper, K., Kolbl, A., Biener, D., Dittrich, S., von Hase, J., Thormeyer, T., Fiegler, H., Carter, N. P., Speicher, M. R., Cremer, T., Cremer, M. (2007) Radial chromatin positioning is shaped by local gene density, not by gene expression. Chromosoma 116, 285–306.PubMedCrossRefGoogle Scholar
  6. 6.
    Maalouf, W. E., Liu, Z., Brochard, V., Renard, J. P., Debey, P., Beaujean, N., Zink, D. (2009) Trichostatin A treatment of cloned mouse embryos improves constitutive heterochromatin remodeling as well as developmental potential to term. BMC Dev Biol 9, 11.PubMedCrossRefGoogle Scholar
  7. 7.
    Werner, M., Wilkens, L., Aubele, M., Nolte, M., Zitzelsberger, H., Komminoth, P. (1997) Interphase cytogenetics in pathology: principles, methods, and applications of fluorescence in situ hybridization (FISH). Histochem Cell Biol 108, 381–390.PubMedCrossRefGoogle Scholar
  8. 8.
    Wassarman, P. M. (2005) Contribution of mouse egg zona pellucida glycoproteins to gamete recognition during fertilization. J Cell Physiol 204, 388–391.PubMedCrossRefGoogle Scholar
  9. 9.
    Bouzinba-Segard, H., Guais, A., Francastel, C. (2006) Accumulation of small murine minor satellite transcripts leads to impaired centromeric architecture and function. Proc Natl Acad Sci U S A 103, 8709–8714.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Walid E. Maalouf
    • 1
    • 2
    Email author
  • Tiphaine Aguirre-Lavin
    • 1
  • Laetitia Herzog
    • 1
  • Isabelle Bataillon
    • 1
  • Pascale Debey
    • 1
  • Nathalie Beaujean
    • 1
  1. 1.UMR 1198 Biologie du Développement et ReproductionINRAJouy en JosasFrance
  2. 2.University of EdinburghEdinburghUK

Personalised recommendations