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The TUNEL assay enables in situ visualization of apoptosis by labeling fragmented DNA, though its effective application requires consideration of recent technical improvements and inherent assay limitations. Novelty: ClaimNovelty.SYNTHESIS. Consensus alignment: ConsensusAlignment.NEUTRAL.

October 15, 2010

In Situ Detection of Apoptosis by the TUNEL Assay: An Overview of Techniques

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Abstract

Apoptosis, or programmed cell death, plays an important role in normal development and homeostasis of adult tissues. Apoptosis has also been linked to many disease states, including cancer. One of the biochemical hallmarks of apoptosis is the generation of free 3'-hydroxyl termini on DNA via cleavage of chromatin into single and multiple oligonuleosome-length fragments. The TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay exploits this biochemical hallmark by labeling the exposed termini of DNA, thereby enabling visualization of nuclei containing fragmented DNA. This review outlines the general method for in situ TUNEL staining of cultured cells and tissue sections, and highlights recent improvements in the technique and limitations of the assay.

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