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NDT402-c Tissue preparation & neurodegeneration detection with in situ nick translation technique (ISNT) in the brain

This service includes sectioning, ISNT labeling, coverslipping and labeling of slides. As a result, you will receive up to 40 ISNT-labeled sections per brain or per tissue block ready for microscopic observations. To express our appreciation, a set of Nissl (or H & E) stained sections adjacent to those used for ISNT labeled sections will also be provided for you without additional cost.

Procedure: Unfixed frozen tissue will be cut on a cryostat and mounted on proteinase-resistant microscope slides (click to see detail). Sections cut through various levels (or the levels of your choice) will be processed on slides for the detection of neurodegeneration with in situ nick translation technique1.

This technique, originally described by Gold et al. (1993)1, labels the nuclei of neurons undergoing DNA fragmentation. This method is based on the ability of DNA polymerase I to catalyze the polymerization of deoxyuridines onto the ends of DNA fragments. The integrated biotins are amplified and visualized by the avidin-biotin-complex methods2 (cf. photo samples below).

1. A quotation is required before placing an order.
2. The investigator needs to provide unfixed frozen tissue.
3. Please contact us for more information.

1. Gold R., Schmied M., Rothe G., Zischler H., Breitschopf H., Wekerle H. and Lassmann H. (1993) Detection of DNA fragmentation in apoptosis: application of in situ nick translation to cell culture systems and tissue sections. J. Histochem. Cytochem. 41: 1023-1030.
2. Hsu S. M., Raine L. and Fanger H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29: 577-580.

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