Harnessing artificial intelligence to reduce phototoxicity in live imaging

Abstract

Fluorescence microscopy is essential for studying living cells, tissues and organisms. However, the fluorescent light that switches on fluorescent molecules also harms the samples, jeopardizing the validity of results – particularly in techniques such as super-resolution microscopy, which demands extended illumination. Artificial intelligence (AI)-enabled software capable of denoising, image restoration, temporal interpolation or cross-modal style transfer has great potential to rescue live imaging data and limit photodamage. Yet we believe the focus should be on maintaining light-induced damage at levels that preserve natural cell behaviour. In this Opinion piece, we argue that a shift in role for AIs is needed – AI should be used to extract rich insights from gentle imaging rather than recover compromised data from harsh illumination. Although AI can enhance imaging, our ultimate goal should be to uncover biological truths, not just retrieve data. It is essential to prioritize minimizing photodamage over merely pushing technical limits. Our approach is aimed towards gentle acquisition and observation of undisturbed living systems, aligning with the essence of live-cell fluorescence microscopy.

PhotoFiTT: A Quantitative Framework for Assessing Phototoxicity in Live-Cell Microscopy Experiments

“…These effects, often subtle and cumulative, can lead to significant changes in cell behaviour, organelle integrity, and developmental processes (6)(7)(8)(9). Yet, establishing optimal imaging protocols that balance high-quality data acquisition with minimal biological interference remains a complex challenge (10). Traditional methods for assessing phototoxicity include viability assays and morphological observations.…”

“…Importantly, tolerance to photodamage varies across specimens and is influenced by damage severity, complicating the development of replicable imaging protocols (12,13). Previous studies have offered valuable quantitative assessments of phototoxicity (3,10,14), but a critical need remains for standardised and generalisable methods able to quantitatively link cell damage to high-intensity light exposure across different fluorescence microscopy techniques. Addressing this gap, we introduce PhotoFiTT (Phototoxicity Fitness Time Trial), a quantitative imaging-based framework designed to assess phototoxicity effects on cellular behaviour in live-cell microscopy experiments (Figure 1).…”