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Histological aspects of in situ hybridization

Detection of poly(A) nucleotide sequences in mouse liver sections as a model system

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Summary

This study examined the detection of cellular poly(A) sequences in mouse liver sections by in situ hybridization using a 3H-labelled poly(dT) probe. Parameters examined included possible losses of target poly(A) sequences from sectioned cells, access of probe to target sequences, section thickness, hybridization conditions, autoradioigraphic efficiency, specific activity of probes and specificity of reaction. An improved protocol was devised that resulted in good preservation of histological detail in sectioned tissue blocks, and a calculated hybridization efficiency of 50%–100%. With the use of probes of defined sequence, the protocol should allow detection of unique mRNA sequences within single cells with an estimated sensitivity of 6–12 unique mRNA molecules per sectioned cell.

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Authors and Affiliations

  1. Division of Medical Virology, Institute of Medical and Veterinary Science, Frome Road, 5000, Adelaide, Australia

    A. R. Jilbert, C. J. Burrell & E. J. Gowans

  2. Tissue Pathology, Institute of Medical and Veterinary Science, Frome Road, 5000, Adelaide, Australia

    R. Rowland

Authors
  1. A. R. Jilbert
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  2. C. J. Burrell
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  3. E. J. Gowans
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  4. R. Rowland
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Jilbert, A.R., Burrell, C.J., Gowans, E.J. et al. Histological aspects of in situ hybridization. Histochemistry 85, 505–514 (1986). https://doi.org/10.1007/BF00508433

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  • DOI: https://doi.org/10.1007/BF00508433

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