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TUNEL assay

2 History
Originally described in the paper by Gavrieli, Sherman,
and Ben-Sasson in 1992,[1] TUNEL has become one of
the main methods for detecting apoptotic programmed
cell death. However, for years there has been a debate
about its accuracy, due to problems in the original assay which caused necrotic cells to be inappropriately labeled as apoptotic.[2] The method has subsequently been
improved dramatically and if performed correctly should
only identify cells in the last phase of apoptosis.[3][4] New
methods incorporate the dUTPs modied by uorophores
or haptens, including biotin or bromine, which can be detected directly in the case of a uorescently-modied nucleotide (i.e., uorescein-dUTP), or indirectly with streptavidin or antibodies, if biotin-dUTP or BrdUTP are
used, respectively. Often at late stages of apoptosis, adherent cells are known to detach or pop o. For a reliMouse liver showing an apoptotic cell stained with TUNEL
able and reproducible TUNEL imaging assay, the modied nucleotide must not only be an acceptable substrate
Terminal deoxynucleotidyl transferase dUTP nick for TdT, but the detection method must also be sensitive
end labeling (TUNEL) is a method for detecting DNA without bringing about any additional loss of cells from
fragmentation by labeling the terminal end of nucleic the sample.
acids.

3 References
[1] Gavrieli Y, Sherman Y, Ben-Sasson SA (1992).
Identication of programmed cell death in situ via
specic labeling of nuclear DNA fragmentation. J Cell
Biol 119 (3): 493501. doi:10.1083/jcb.119.3.493.
PMC 2289665. PMID 1400587.
[2] Grasl-Kraupp B, Ruttkay-Nedecky B, Koudelka H,
Bukowska K, Bursch W, Schulte-Hermann R (1995). In
situ detection of fragmented DNA (TUNEL assay) fails to
discriminate among apoptosis, necrosis, and autolytic cell
death: a cautionary note. Hepatology 21 (5): 14658.
doi:10.1002/hep.1840210534. PMID 7737654.

dUTP

[3] Negoescu A, Lorimier P, Labat-Moleur F, Drouet C,


Robert C, Guillermet C, Brambilla C, Brambilla E
(1996). In situ apoptotic cell labeling by the TUNEL
method: improvement and evaluation on cell preparations.
J Histochem Cytochem 44 (9): 95968.
doi:10.1177/44.9.8773561. PMID 8773561.

Method

TUNEL is a common method for detecting DNA fragmentation that results from apoptotic signaling cascades.
The assay relies on the presence of nicks in the DNA
which can be identied by terminal deoxynucleotidyl
transferase or TdT, an enzyme that will catalyze the addition of dUTPs that are secondarily labeled with a marker.
It may also label cells that have suered severe DNA
damage.

[4] Negoescu A, Guillermet C, Lorimier P, Brambilla


E, Labat-Moleur F (1998). Importance of DNA
fragmentation in apoptosis with regard to TUNEL
specicity. Biomed Pharmacother 52 (6): 2528.
doi:10.1016/S0753-3322(98)80010-3. PMID 9755824.

External links
TUNEL at the US National Library of Medicine
Medical Subject Headings (MeSH)

EXTERNAL LINKS

Text and image sources, contributors, and licenses

5.1

Text

TUNEL assay Source: http://en.wikipedia.org/wiki/TUNEL_assay?oldid=645636490 Contributors: Nyh, Arcadian, Pearle, Conny,


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5.2

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File:Apoptosis_stained.jpg Source: https://upload.wikimedia.org/wikipedia/commons/5/5f/Apoptosis_stained.jpg License: Public domain Contributors: http://dir.niehs.nih.gov/dirlep/liverpath/necrotic/apoptosis5.htm Original artist: Laboratory of Experimental Pathology,
Division of Intramural Research, NIEHS (NIH)
File:DNA_stub.png Source: https://upload.wikimedia.org/wikipedia/commons/6/61/DNA_stub.png License: CC-BY-SA-3.0 Contributors: Transferred from en.wikipedia; transfer was stated to be made by User:Rockfang. Original artist: Original uploader was Alai at
en.wikipedia
File:DUTP_chemical_structure.png Source: https://upload.wikimedia.org/wikipedia/commons/f/fb/DUTP_chemical_structure.png
License: CC-BY-SA-3.0 Contributors: ? Original artist: ?

5.3

Content license

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