Active and Probe-Free Intracellular Rheology via Phase-Sensitive Thermoviscous Flows

Determination of the rheological properties of cells is known to require active measurements, which largely depend on the internalisation of mechanical probes. Here we circumvent this problem via the introduction of Rheo-FLUCS: an active, yet probe-free approach that leverages light-induced flows to access the mechanics of complex systems. While Rheo-FLUCS is facilitated by thermoviscous expansion phenomena rather than external forces, we here show equivalence in its ability to measure canonical viscoelastic properties. Specifically, we demonstrate a phase-lag equivalence with probe-dependent active microrheology in a wide range of physically different, yet chemically identical materials. We exemplify the utility of Rheo-FLUCS in three distinctly different biological systems: compound-treated mouse fibroblasts (NIH-3T3), genetically modified human osteoblasts (U2OS) to elucidate the role of myosins in cytoplasmic mechanics, and early ascidian oocytes of Phallusia mammillata at fertilisation stage. This probe-free methodology represents a transformative advance in cellular rheology, enabling non-invasive and precise mechanical measurements across diverse biological systems without perturbing their natural state through probe internalisation.