Description

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.

Procedure:  Unfixed frozen tissue will be cut on a cryostat and mounted on proteinase-resistant microscope slides (cf. Products, Cat. #PO102). 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 situDNA 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 (cf. photo samples).

Remarks:

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

References:

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.