CometAnalyser: a user-friendly, open-source deep-learning microscopy tool for quantitative comet assay analysis

Comet assay provides an easy solution to estimate DNA damage in single cells through microscopy assessment. It is widely used in the analysis of genotoxic damages induced by radiotherapy or chemotherapeutic agents. DNA damage is quantified at the single-cell level by computing the displacement between the genetic material within the nucleus, typically called “comet head”, and the genetic material in the surrounding part of the cell, considered as the “comet tail”. Today, the number of works based on Comet Assay analyses is really impressive. In this work, besides revising the solutions available to obtain reproducible and reliable quantitative data, we developed an easy-to-use tool named CometAnalyser. It is designed for the analysis of both fluorescent and silver-stained wide-field microscopy images and allows to automatically segment and classify the comets, besides extracting Tail Moment and several other intensity/morphological features for performing statistical analysis. CometAnalyser is an open-source deep-learning tool. It works with Windows, Macintosh, and UNIX-based systems. Source code, standalone versions, user manual, sample images, video tutorial and further documentation are freely available at: https://sourceforge.net/p/cometanalyser.HIGHLIGHTSComet assay provides an easy solution to estimate DNA damage in single cells.Today, an impressive number of works are based on Comet Assay analyses, especially in the field of cancer research.Comet assay was originally performed as a qualitative analysis.None of the free tools today available work on both fluorescent- and silver-stained images.We developed CometAnalyser, an open-source deep-learning tool designed for easy segmentation and classification of comets in fluorescent- and silver-stained images.