DNA损伤是癌症和神经退行性疾病中常见的细胞特征，但快速而准确的DNA损伤定量方法缺乏。彗星实验是一种基于断裂的DNA链向正极迁移而创建细胞后有可量化的“尾巴”的生化工具，用于测量DNA损伤。然而，这种方法的主要限制是需要对可用的开源算法的图像中的彗星进行分析的时间。需要手动审核以及艰辛的前期和后期处理步骤可能需要耗费几小时到几天的时间。为了克服这些限制，我们开发了AutoComet，一种新的开源算法，用于彗星分析，利用自动彗星分割和尾部参数的量化。AutoComet首先根据大小和强度分段和过滤彗星，然后过滤掉没有良好连接的头和尾的彗星，这显着增加了分割精度。由于AutoComet是完全自动化的，因此可以最小化审核员的偏差，可扩展，将分析时间减少了十倍以上，减少了每个彗星不到3秒的时间。AutoComet成功地检测到了有诱导DNA损伤与没有DNA损伤的细胞之间尾部参数的显着差异，并且与手动审核的结果相比，比其他开源软件分析程序更加可比。我们得出的结论是，AutoComet算法提供了一种快速、无偏差和准确量化DNA损伤的方法，避免了手动审核的固有限制，并将显着提高检测DNA损伤的能力。版权所有©2023 Elsevier B.V. 发布。

DNA damage is a common cellular feature seen in cancer and neurodegenerative disease, but fast and accurate methods for quantifying DNA damage are lacking. Comet assays are a biochemical tool to measure DNA damage based on the migration of broken DNA strands towards a positive electrode, which creates a quantifiable 'tail' behind the cell. However, a major limitation of this approach is the time needed for analysis of comets in the images with available open-source algorithms. The requirement for manual curation and the laborious pre- and post-processing steps can take hours to days. To overcome these limitations, we developed AutoComet, a new open-source algorithm for comet analysis that utilizes automated comet segmentation and quantification of tail parameters. AutoComet first segments and filters comets based on size and intensity and then filters out comets without a well-connected head and tail, which significantly increases segmentation accuracy. Because AutoComet is fully automated, it minimizes curator bias and is scalable, decreasing analysis time over ten-fold, to less than 3 s per comet. AutoComet successfully detected statistically significant differences in tail parameters between cells with and without induced DNA damage, and was more comparable to the results of manual curation than other open-source software analysis programs. We conclude that the AutoComet algorithm provides a fast, unbiased and accurate method to quantify DNA damage that avoids the inherent limitations of manual curation and will significantly improve the ability to detect DNA damage.Copyright © 2023. Published by Elsevier B.V.