2024/07/16 by Mario Del Rosario, Estibaliz Gómez‐de‐Mariscal, Leonor Morgado +4 · 1 voice
Engineering · Biochemistry, Genetics and Molecular Biology · Veterinary · #3D Printing in Biomedical Research #bioluminescence and chemiluminescence research #Animal testing and alternatives
paper · doi:10.1101/2024.07.16.603046
openalex publication_date 2024/07/16 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/01
Phototoxicity in live-cell fluorescence microscopy can compromise experimental outcomes, yet quantitative methods to assess its impact remain limited. Here we present PhotoFiTT (Phototoxicity Fitness Time Trial), an integrated framework combining a standardised experimental protocol with advanced image analysis to quantify light-induced cellular stress in label-free settings. PhotoFiTT leverages machine learning and cell cycle dynamics to analyse mitotic timing, cell size changes, and overall cellular activity in response to controlled light exposure. Using adherent mammalian cells, we demonstrate PhotoFiTT’s ability to detect wavelength- and dose-dependent effects, showcasing that near-UV light induces significant mitotic delays at doses as low as 0.6 J/cm 2 , while longer wavelengths require higher doses for comparable effects. PhotoFiTT enables researchers to establish quantitative benchmarks for acceptable levels of photodamage, facilitating the optimisation of imaging protocols that balance image quality with sample health.