In Situ Hybridization

In situ hybridization :

The techniques involves the detection of specific nucleic acid sequences (DNA or RNA) inside the cell , using specific probes.

 

 

EBER Probe:

The probe is used for the identification of latent infection caused by the Epstein –Barr virus (EBV) associated with several diseases, such Hodgkin’s lymphoma, B cell lymphoma, nasopharyngeal carcinoma  and others. 

 

Kappa and Lambda Probes: 

Kappa and Lambda probes are used for identification of mRNA of kappa and lambda light chains from the immunoglobulins of B cells.

 

PNA ISH Detection Kit:

Is the method used in SQUH and the kit intended for the detection of fluorescein-conjugated peptide nucleic acid (PNA) probes hybridized to their target RNA in cell or tissue preparation.

 

EBER, Kappa and Lambda (mRNA) PNA Probes are labeled with fluorescein and used with PNA ISH Detection kit from Dako and visualized by substrate ( enzyme substrate , BCIP/NBT) and the positive staining is recognized as a dark blue/black color at the site of hybridization.

Silver In Situ Hybridization

Detection of Her2/neu using SISH Method:

 

The Her2 gene is amplified and over-expressed in forms of breast cancer, ovarian cancer, endometrial cancer, gastric cancer and others. It represents a predictive factor for therapeutic assessment of patients for whom Herceptin (Trastuzumab) treatment is  being considered.

 

Her2 Probe: INFORM HER2 Dual ISH DNA Probe Cocktail(Ventana)

 

The INFORM HER2 Dual ISH DNA Probe Cocktail is formulated for use with Ventana ultraView SISH DNP detection kit and Ventana ultraView Red ISH DIG detection kit on Ventana Automated Stainer.

 

In the SISH method , the HER2 gene is detected by hybridization with a labeled DNA probe  with Dinitrophenol (DNP) and visualized with silver , followed by hybridization with the chromosome 17, labeled  AP dephosphorylate naththol and visualized Fast red. The results are visualized by light microscope