Systematic comparison of genetically encoded F-actin probes for visualizing actomyosin dynamics in vivo using C. elegans

In Caenorhabditis elegans, mStayGold–F-actin probes delivered the highest brightness and photostability, supporting long-term time-lapse imaging across multiple tissues and developmental stages.

• Why it matters: Probe choice can change both imaging quality and cellular behavior, yet performance of genetically encoded F-actin reporters is context-dependent and not well standardized for in vivo use.
• What they did: The study compared F-tractin, LifeAct, and Utrophin fused to mScarlet-I, mNeonGreen, or mStayGold in single-copy transgenic C. elegans, measuring fluorescence intensity, photostability, developmental and reproductive outcomes, cytokinetic ring closure, and localization across cellular contexts.
• The result: mNeonGreen provided an alternative where photobleaching is acceptable, most strains showed minimal detectable effects and improved over previously generated lines, and probe performance varied by tissue/stage with F-tractin showing lower embryo fluorescence.

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