PhotoFiTT: a quantitative framework for assessing phototoxicity in live-cell microscopy experiments.

ABSTRACT: 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/cm², while longer wavel

SUBMITTER: Del Rosario M 

PROVIDER: S-EPMC12738901 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature