TScratch: a novel and simple software tool for automated analysis of monolayer wound healing assays

TLDR

Cell migration is essential in development, physiology, and disease and is routinely assessed with monolayer wound‑healing assays, yet manual analysis is laborious. TScratch was created to automate this assay by applying a fast discrete curvelet transform to quantify cell‑occupied area in images. The software offers a graphical interface for inspecting and adjusting results and was validated against manual measurements across several cell lines. TScratch markedly shortens analysis time, delivers objective and reproducible quantification, and closely matches manual results, reliably reproducing significant effects of known migration inhibitors and enhancers.

Abstract

Cell migration plays a major role in development, physiology, and disease, and is frequently evaluated in vitro by the monolayer wound healing assay. The assay analysis, however, is a time-consuming task that is often performed manually. In order to accelerate this analysis, we have developed TScratch, a new, freely available image analysis technique and associated software tool that uses the fast discrete curvelet transform to automate the measurement of the area occupied by cells in the images. This tool helps to significantly reduce the time needed for analysis and enables objective and reproducible quantification of assays. The software also offers a graphical user interface which allows easy inspection of analysis results and, if desired, manual modification of analysis parameters. The automated analysis was validated by comparing its results with manual-analysis results for a range of different cell lines. The comparisons demonstrate a close agreement for the vast majority of images that were examined and indicate that the present computational tool can reproduce statistically significant results in experiments with well-known cell migration inhibitors and enhancers.