In Situ Hybridization (ISH)

Introduction

In Situ Hybridization (ISH)

is a technique that allows for precise localization of a specific segment of nucleic acid within a histologic section. The underlying basis of ISH is that nucleic acids, if preserved adequately within a histologic specimen, can be detected through the application of a complementary strand of nucleic acid to which a reporter molecule is attached.

Visualization of the reporter molecule allows to localize DNA or RNA sequences in a heterogeneous cell populations including tissue samples and environmental samples. Riboprobes also allow to localize and assess degree of gene expression . The technique is particularly useful in neuroscience.

In situ hybridization probes

  • Double-stranded DNA (dsDNA) probes
  • Single-stranded DNA (ssDNA) probes
  • RNA probes (riboprobes)
  • Synthetic oligonucleotides (PNA, LNA)

Labeling techniques

  • Radioactive isotopes
    • 32P
    • 35S
    • 3H
  • Non-radioactive labels
    • biotin
    • digoxigenin
    • fluorescent dye (FISH)

Applications of In Situ Hybridization

  • Microbiology (classic target - 16S rRNA) - morphology and population structure of microorganisms
  • Pathology (pathogen profiling, abnormal gene expression)
  • Developmental biology (gene expression profiling in embryonic tissues)
  • Karyotyping and phylogenetic analysis (unique FISH patterns on individual chromosomes, chromosomal aberrations)
  • Physical mapping (mapping clones on chromosomes and direct assignment of mapped clones to chromosomal regions associated with heterochromatin or euchromatin)

Generation of riboprobes

Generation of riboprobes

References

Note: [MAJR] is a Medical Subject Heading (MeSH) tag for Major Heading. The tag is used to limit the search to articles for which major subjects are represented by terms included in the NLM MeSH database.

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Last updated: 2014-09-24T14:00:35-04:00