Skip to main content

Preparation of Tissue Sections and Slides for mRNA Hybridization

  • Protocol
Book cover Protocols for Nucleic Acid Analysis by Nonradioactive Probes

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 28))

  • 673 Accesses

Abstract

The first step for a successful in situ hybridization is the fixation of the tissue. This will ensure target nucleic acid retention and preservation of the tissue morphology. Either crosslinking or precipitative fixatives can be used, and a preference for either of the two types of fixative has often been based on the different types of system under investigation (17). For hybridization of regulatory peptide mRNA, 4% paraformaldehyde appears to be the most effective, both on tissue blocks and on tissue culture preparations.

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 74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Haase, A. T., Brahic, M., and Stowring, L. (1984) Detection of viral nucleic acids by in situ hybridization, in Methods in Virology, vol. VII (Maramorosch, K. and Koprowski, H., eds.), Academic, New York, pp. 189–226.

    Google Scholar 

  2. McAllister, H. A. and Rock, D. L. (1985) Comparative usefulness of tissue fixatives for in situ viral nucleic acid hybridization. J. Histochem. Cytochem. 33, 1026–1032.

    PubMed  CAS  Google Scholar 

  3. Moench, T. R., Gendelman, H. E., Clements, J. E., Narayan, O., and Griffin, D. E. (1985) Efficiency of in situ hybridization as a function of probe size and fixation technique. J. Virol. Method. 11, 119–130.

    Article  CAS  Google Scholar 

  4. Hofler, H., Childers, H., Montminy, M. R., Lechan, R. M., Goodman, R. H., and Wolfe, H. J. (1986) In situ hybridization methods for the detection of somatostatin mRNA in tissue sections using antisense RNA probes. Histochem. J. 18, 597–604.

    Article  Google Scholar 

  5. Singer, R. H., Lawrence, J. B., and Villnave, C. (1986) Optimization of in situ hybridization using isotopic and non-isotopic detection methods. Biotechniques 4, 230–250.

    CAS  Google Scholar 

  6. Guitteny, A. F., Fouque, B., Mongin, C., Teoule, R., and Boch, B. (1988) Histological detection of mRNA with biotinylated synthetic oligonucleotide probes. J. Histochem. Cytochem. 36, 563–571.

    PubMed  CAS  Google Scholar 

  7. Terenghi, G. and Fallon, R. A. (1990) Techniques and applications of in situ hybridization, in Current Topics in Pathology: Pathology of the Nucleus (Underwood, J. C. E., ed.), Springer Verlag, Berlin, pp. 290–337.

    Google Scholar 

  8. Farquharson, M., Harvie, R., and McNicol, A. M. (1990) Detection of mRNA using a digoxigenin end labelled oligodeoxynucleotide probe. J. Clin. Pathol. 43, 424–428.

    Article  PubMed  CAS  Google Scholar 

  9. Unger, E. R., Hammer, M. I., and Chenggis, M. L. (1991) Comparison of 35S and biotin as labels for in situ hybridization: use of an HPV model system. J. Histochem. Cytochem. 39, 145–150.

    PubMed  CAS  Google Scholar 

  10. Asanuma, M., Ogawa, N., Mizukawa, K., Haba, K., and Mori, A. (1990) A comparison of formaldehyde-preperfused frozen and freshly frozen tissue preparation for the in situ hybridization for α-tubulin mRNA in the rat brain. Res. Comm. Chem. Pathol. Pharmacol. 70, 183–192.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1994 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Terenghi, G., Polak, J.M. (1994). Preparation of Tissue Sections and Slides for mRNA Hybridization. In: Isaac, P.G. (eds) Protocols for Nucleic Acid Analysis by Nonradioactive Probes. Methods in Molecular Biology™, vol 28. Humana Press. https://doi.org/10.1385/0-89603-254-X:187

Download citation

  • DOI: https://doi.org/10.1385/0-89603-254-X:187

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-254-5

  • Online ISBN: 978-1-59259-515-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics