In Situ Hybridization to Sections (Nonradioactive)

  • Maria Rex
  • Paul J. Scotting
Part of the Methods in Molecular Biology™ book series (MIMB, volume 97)


In situ hybridization (ISH) takes advantage of the ability of mRNA within a cell to hybridize with exogenously applied complementary RNA (riboprobes) or DNA molecules. This interaction is visualized by labeling the applied nucleic acid probe with a detectable molecule (radioactive, such as 35S, or non-radioactive, such as digoxygenin [DIG]). The technique allows patterns of gene expression to be visualized in many tissues or cell types simultaneously.


Color Detection Sheep Serum Blue Tetrazolium Chloride Alkaline Phosphatase Streptavidin Fume Cupboard 
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  1. 1.
    Rex, M. and Scotting, P. J. (1994) Simultaneous detection of RNA and protein in tissue sections by nonradioactive in situ hybridization followed by immunohistochemistry. Biochemica (Boehringer Mannheim) 3, 24–26.Google Scholar
  2. 2.
    Dickerson, D. S., Huerter, B. S., Morris, S. J., and Chronwall, B. M. (1994) POMC mRNA levels in individual melanotropes and GFAP in glial-like cells in rat pituitary. Peptides 15, 247–256.PubMedCrossRefGoogle Scholar
  3. 3.
    Wang, D. and Cutz, E. (1994) Simultaneous detection of mRNA for bombesin/gastrin-releasing peptide and its receptor in rat brain by nonradiolabeled double ISH. Laboratory Investigation 70, 775–780.PubMedGoogle Scholar
  4. 4.
    Angerer, L and Angerer, R. (1991) Localisation of mRNAs by in situ hybridization. Meth. Cell Biol. 35, 37–71.CrossRefGoogle Scholar
  5. 5.
    Norton, A.J., Jordan, S., and Yeomans, P. (1994) Brief, high temperature heat denaturation (pressure cooking): a simple and effective method of antigen retrieval for routinely processed tissues. J. Pathol. 173, 371–379.PubMedCrossRefGoogle Scholar
  6. 6.
    Brouwer, N., Van Dijken, H., Ruiters, M. H. J., Van Willigen, J.-D., and Horst, G. J. (1992) Localisation of dopamine D2 receptor mRNA with non-radioactive in situ hybridization histochemistry. Neurosci. Lett. 142, 223–227.PubMedCrossRefGoogle Scholar
  7. 7.
    Komminoth, P., Merk, F. B., Leav, I., Wolfe, H. J., and Roth, J. (1992) Comparison of 35S-and digoxigenin-labeled RNA and oligonucleotide probes for ISH. Histochemistry 98, 217–228.PubMedCrossRefGoogle Scholar
  8. 8.
    Rex, M. Uwanogho, D. A., Orme, A., Scotting, P. J., and Sharpe, P. T. (1997) cSox21 exhibits a complex and dynamic pattern of transcription during embryonic development of the chick central nervous system. Med. Dev. 66, 39–53.Google Scholar
  9. 9.
    Uwanogho, D., Rex, M., Cartwright, E., Pearl, G., Scotting, P. J., and Sharpe, P. T. (1995) Embryonic expression of the chicken Sox2, Sox3 and Sox11 genes suggests an interactive role in neuronal development. Mech. Dev. 49, 23–36.PubMedCrossRefGoogle Scholar
  10. 10.
    Rex, M., Church, R., Tointon, K., and Scotting, P. J. (1997) Combination of non-isotopic in situ hybridization with detection of enzyme activity, bromodeoxyuridine incorporation and immunohistochemical markers. Histochem. Cell Biol. 107, 519–523.PubMedCrossRefGoogle Scholar
  11. 11.
    Rex, M., Orme, A., Uwanogho, D., Tointon, K., Wigmore, P. M., Sharpe, P. T., and Scotting, P. J. (1997) Dynamic expression of chicken Sox2 and Sox3 genes in ectoderm induced to form neural tissue. Dev. Dynamics 209, 323–332.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1999

Authors and Affiliations

  • Maria Rex
    • 1
  • Paul J. Scotting
    • 1
  1. 1.Department of GeneticsUniversity of NotinghamUK

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