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Getting a grip on how we may age healthily is a central interest of biogerontological research. To this end, a number of academic teams developed platforms for life-and healthspan assessment in Caenorhabditis elegans. These are very appealing for medium-to high throughput screens, but a broader implementation is lacking due to many systems relying on custom scripts for data analysis that others struggle to adopt. Hence, user-friendly recommendations would help to translate raw data into interpretable results. The aim of this communication is to streamline the analysis of data obtained by the WorMotel, an economically and practically appealing screening platform, in order to facilitate the use of this system by interested researchers. We here detail recommendations for the stepwise conversion of raw image data into activity values and explain criteria for assessment of health in C. elegans based on locomotion. Our analysis protocol can easily be adopted by researchers, and all needed scripts and a tutorial are available in S1 and S2 Files. OPEN ACCESS Citation: Jushaj A, Churgin M, Yao B, De La Torre M, Fang-Yen C, Temmerman L (2020) Optimized criteria for locomotion-based healthspan evaluation in C. elegans using the WorMotel system. PLoS ONE 15(3): e0229583. https://doi.org/10.

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