biostudies-literatureEbata HJapan Society for the Promotion of Science1781-1793https://www.ebi.ac.uk/biostudies/studies/S-EPMC10209042biostudies-literatureUnknown122(10)Active microrheology was conducted in living cells by applying an optical-trapping force to vigorously fluctuating tracer beads with feedback-tracking technology. The complex shear modulus G(ω)=G^'(ω)-iG^″(ω) was measured in HeLa cells in an epithelial-like confluent monolayer. We found that G(ω)∝(-iω)^1/2 over a wide range of frequencies (1 Hz < ω/2π < 10 kHz). Actin disruption and cell-cycle progression from G1 to S and G2 phases only had a limited effect on G(ω) in living cells. On the other hand, G(ω) was found to be dependent on cell metabolism; ATP-depleted cells showed an increased elastic modulus G^'(ω) at low frequencies, giving rise to a constant plateau such that G(ω)=G₀+A(-iω)^1/2. Both the plateau and the additional frequency dependency ∝(-iω)^1/2 of ATP-depleted cells are consistent with a rheological response typical of colloidal jamming. On the other hand, the plateau G₀ disappeared in ordinary metabolically active cells, implying that living cells fluidize their internal states such that they approach the critical jamming point.Biophysical journalActivity-dependent glassy cell mechanics Ⅰ: Mechanical properties measured with active microrheology.PMC10209042JP21H01048JP20H05536JP20H00128JP22H04848Mizuno DInokuchi SMiyamoto TNishizawa KUmeda KSugino YEbata HNagao WfalseActivity-dependent glassy cell mechanics Ⅰ: Mechanical properties measured with active microrheology.Active microrheology was conducted in living cells by applying an optical-trapping force to vigorously fluctuating tracer beads with feedback-tracking technology. The complex shear modulus G(ω)=G^'(ω)-iG^″(ω) was measured in HeLa cells in an epithelial-like confluent monolayer. We found that G(ω)∝(-iω)^1/2 over a wide range of frequencies (1 Hz < ω/2π < 10 kHz). Actin disruption and cell-cycle progression from G1 to S and G2 phases only had a limited effect on G(ω) in living cells. On the other hand, G(ω) was found to be dependent on cell metabolism; ATP-depleted cells showed an increased elastic modulus G^'(ω) at low frequencies, giving rise to a constant plateau such that G(ω)=G₀+A(-iω)^1/2. Both the plateau and the additional frequency dependency ∝(-iω)^1/2 of ATP-depleted cells are consistent with a rheological response typical of colloidal jamming. On the other hand, the plateau G₀ disappeared in ordinary metabolically active cells, implying that living cells fluidize their internal states such that they approach the critical jamming point.2023-01-01T00:00:00Z2023 May2025-06-27T03:05:21.551Z2025-06-27T03:05:21.551ZS-EPMC102090423705087510.1016/j.bpj.2023.04.011