Visual comet scoring revisited: a guide to scoring comet assay slides and obtaining reliable results
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Visual comet scoring revisited : a guide to scoring comet assay slides and obtaining reliable results. / Møller, Peter; Azqueta, Amaya; Sanz-Serrano, Julen; Bakuradze, Tamara; Richling, Elke; Bankoglu, Ezgi Eyluel; Stopper, Helga; Bastos, Victoria Claudino; Langie, Sabine A S; Jensen, Annie; Scavone, Francesca; Giovannelli, Lisa; Wojewódzka, Maria; Kruszewski, Marcin; Valdiglesias, Vanessa; Laffon, Blanca; Costa, Carla Trindade; Costa, Solange; Teixeira, João Paulo; Marino, Mirko; Del Bo', Cristian; Riso, Patrizia; Zheng, Congying; Shaposhnikov, Sergey; Collins, Andrew.
In: Mutagenesis, Vol. 38, No. 5, 2023, p. 253–263.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Visual comet scoring revisited
T2 - a guide to scoring comet assay slides and obtaining reliable results
AU - Møller, Peter
AU - Azqueta, Amaya
AU - Sanz-Serrano, Julen
AU - Bakuradze, Tamara
AU - Richling, Elke
AU - Bankoglu, Ezgi Eyluel
AU - Stopper, Helga
AU - Bastos, Victoria Claudino
AU - Langie, Sabine A S
AU - Jensen, Annie
AU - Scavone, Francesca
AU - Giovannelli, Lisa
AU - Wojewódzka, Maria
AU - Kruszewski, Marcin
AU - Valdiglesias, Vanessa
AU - Laffon, Blanca
AU - Costa, Carla Trindade
AU - Costa, Solange
AU - Teixeira, João Paulo
AU - Marino, Mirko
AU - Del Bo', Cristian
AU - Riso, Patrizia
AU - Zheng, Congying
AU - Shaposhnikov, Sergey
AU - Collins, Andrew
N1 - © The Author(s) 2023. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
PY - 2023
Y1 - 2023
N2 - Measurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20-25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 different laboratories scored the comet images using a five-class scoring system. There is inter-investigator variation in the three training sets of comets (i.e. coefficient of variation (CV) = 9.7%, 19.8% and 15.2% in training sets I-III, respectively). However, there is also a positive correlation of inter-investigator scoring in the three training sets (r = 0.60). Overall, 36% of the variation is attributed to inter-investigator variation and 64% stems from intra-investigator variation in scoring between comets (i.e. the comets in training sets I-III look slightly different and this gives rise to heterogeneity in scoring). Intra-investigator variation in scoring was also assessed by repeated analysis of the training sets by the same investigator. There was larger variation when the training sets were scored over a period of six months (CV = 5.9-9.6%) as compared to one week (CV = 1.3-6.1%). A subsequent study revealed a high inter-investigator variation when premade slides, prepared in a central laboratory, were stained and scored by investigators in different laboratories (CV = 105% and 18-20% in premade slides with comets from unexposed and hydrogen peroxide-exposed cells, respectively). The results indicate that further standardization of visual scoring is desirable. Nevertheless, the analysis demonstrates that visual scoring is a reliable way of analysing DNA migration in comets.
AB - Measurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20-25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 different laboratories scored the comet images using a five-class scoring system. There is inter-investigator variation in the three training sets of comets (i.e. coefficient of variation (CV) = 9.7%, 19.8% and 15.2% in training sets I-III, respectively). However, there is also a positive correlation of inter-investigator scoring in the three training sets (r = 0.60). Overall, 36% of the variation is attributed to inter-investigator variation and 64% stems from intra-investigator variation in scoring between comets (i.e. the comets in training sets I-III look slightly different and this gives rise to heterogeneity in scoring). Intra-investigator variation in scoring was also assessed by repeated analysis of the training sets by the same investigator. There was larger variation when the training sets were scored over a period of six months (CV = 5.9-9.6%) as compared to one week (CV = 1.3-6.1%). A subsequent study revealed a high inter-investigator variation when premade slides, prepared in a central laboratory, were stained and scored by investigators in different laboratories (CV = 105% and 18-20% in premade slides with comets from unexposed and hydrogen peroxide-exposed cells, respectively). The results indicate that further standardization of visual scoring is desirable. Nevertheless, the analysis demonstrates that visual scoring is a reliable way of analysing DNA migration in comets.
U2 - 10.1093/mutage/gead015
DO - 10.1093/mutage/gead015
M3 - Journal article
C2 - 37233347
VL - 38
SP - 253
EP - 263
JO - Mutagenesis
JF - Mutagenesis
SN - 0267-8357
IS - 5
ER -
ID: 357731643