Skip to main content

Fluorescence in situ Hybridization (FISH) on Tissue Cryosections

  • Protocol
  • First Online:
Fluorescence in situ Hybridization (FISH)

Part of the book series: Methods in Molecular Biology ((MIMB,volume 659))

Abstract

Recent progress in the understanding of the spatial organization of nuclear functions owes a lot to fluorescence in situ hybridization (FISH) methodology. The majority of studies using this technology have been carried out using cultured cells. However, nuclear processes in whole organisms, may be to a notable degree, different from those in cultured cells and actually not similar across different tissues. Therefore, for better understanding of nuclear processes in ex vivo organismal material, it is necessary to study nuclear organization in sections of tissue. FISH on sections is still not common in nuclear biology studies mostly due to methodological problems. The protocol suggested in this chapter is based on several years experience in hybridizing different probes on cryosections of various tissues.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Cremer, M., Grasser, F., Lanctot, C., Müller, S., Neußer, M., Zinner, R., Solovei, I., and Cremer, T. (2008) Multicolor 3D Fluorescence In Situ Hybridization for Imaging Interphase Chromosomes, in Methods in Molecular Biology: The Nucleus (Hancock, R., Ed.) pp 205–239, Humana Press.

    Google Scholar 

  2. Cremer, M., Müller, S., Köhler, D., Brero, A., and Solovei, I. (2007) in CSH Protocols pp doi:10.1101/pdb.prot4723.

  3. Solovei, I., and Cremer, M. (2010) Combination of Immunostaining and 3D-FISH on Cultured Cells, in Methods in Molecular Biology: FISH (Bridger J. M., V. E., Ed.) Humana Press.

    Google Scholar 

  4. Solovei, I., Walter, J., Cremer, M., Habermann, H., Schermelleh, L., and Cremer, T. (2002) FISH on Three-Dimensionally Preserved Nuclei, in FISH (Beatty B., Larson S. M., J. Squire, Ed.) pp 119–157, Oxford University Press, Oxford.

    Google Scholar 

  5. Solovei, I., Grasser, F., and Lanctôt, C. (2007) in CSH Protocols pp doi:10.1101/pdb.prot4729.

  6. Shi, S. R., Cote, R. J., and Taylor, C. R. (1997) Antigen retrieval immunohistochemistry: past, present, and future. J Histochem Cytochem 45, 327–43.

    Article  PubMed  CAS  Google Scholar 

  7. Shi, S. R., Cote, R. J., and Taylor, C. R. (2001) Antigen retrieval techniques: current perspectives. J Histochem Cytochem 49, 931–7.

    Article  PubMed  CAS  Google Scholar 

  8. Henegariu, O., Bray-Ward, P., and Ward, D. C. (2000) Custom fluorescent-nucleotide synthesis as an alternative method for nucleic acid labeling. Nat Biotechnol 18, 345–8.

    Article  PubMed  CAS  Google Scholar 

  9. Müller, S., Neusser, M., Köhler, D., and Cremer, M. (2007) in CSH Protocols pp doi:10.1101/pdb.prot4730.

  10. Nimmakayalu, M., Henegariu, O., Ward, D. C., and Bray-Ward, P. (2000) Simple method for preparation of fluor/hapten-labeled dUTP. Biotechniques 28, 518–22.

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The author thanks Yana Feodorova and Süleyman Kösem who, as graduate students, carried out some of the immuno-FISH experiments described in this protocol and Mike Fessing (University of Bradford) for providing BAC DNA for Rps27. This work was supported by DFG grant SO1054/1.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Irina Solovei .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Solovei, I. (2010). Fluorescence in situ Hybridization (FISH) on Tissue Cryosections. In: Bridger, J., Volpi, E. (eds) Fluorescence in situ Hybridization (FISH). Methods in Molecular Biology, vol 659. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-789-1_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-60761-789-1_5

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-788-4

  • Online ISBN: 978-1-60761-789-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics