This service includes sectioning, TUNEL labeling, coverslipping and labeling of slides. As a result, you will receive up to 40 TUNEL-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 TUNEL-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 (see Products). Sections cut through various levels (or the levels of your choice) will then be processed on slides for the detection of neurons undergoing apoptosis with in situ DNA nick end-labeling1.
This technique, originally described by Gavrieli et al.(1992)1, labels the nuclei of neurons undergoing DNA fragmentation. This method is based on the ability of terminal deoxynucleotidyl transferase to catalyze incorporation of biotinylated deoxyuridines onto the free 3'-hydroxyl termini of DNA fragments, which are considered as one of the most characteristic features of apoptosis2, 3. The integrated biotins are amplified and visualized by the avidin-biotin-complex (ABC) method4 (see NDT102).
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. Gavrieli Y, Sherman Y, and Ben-Sasson SA: Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119: 493-501, 1992.
2. Wyllie A. H. (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555-556.
3. Arends M. J., Morris R. G. and Wyllie A. H. (1990) Apoptosis: the role of the endonuclease. Amer. J. Pathol. 136: 593-608.
4. 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.