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In situ hybridization
- 1. Presented by Neelma Nayab Biotechnology and GE Mphil 2nd semester kust
- 2. In situ hybridization • In situ hybridization (ISH) is a powerful technique for localizing specific nucleic acid targets within fixed tissues and cells, allowing you to obtain information about gene expression and genetic loci.
- 3. Type of ISH Fluorescence (FISH) Chromogenic (CISH) detection
- 4. Fluorescence in situ hybridization • fluorescence in situ hybridization is a molecular cytogenic technique that uses fluorescence probes that binds to only those parts of the chromosomes with a high degree of sequence complementarity. • It is used to detect and localize the presence or absence of specific D NA sequences on chromosomes.
- 6. Chromogenic in situ hybridization • Chromogenic in situ hybridization (CISH) enables you to gain genetic information in the context of tissue morphology
- 8. Probes • A probe is defined as a single stranded piece of DNA, labelled (either with radioisotope or with non-radioactive label), the nucleotide sequence of which is complementary to the target DNA
Editor's Notes
- Conceptual schematic of single-colour chromogenic in situ hybridisation demonstrating bright field detection of a digoxigenin-labelled probe. The probe is recognised by an antidigoxigenin fluorescein isothiocyanate primary antibody followed by detection with an anti-fluorescein-isothiocyanate horseradish peroxidase (HRP). After addition and oxidation of diaminobenzidine, a dark brown signal is deposited at the target site.