Products Cited in Publications Worldwide Real-time nanomechanical visualization and profiling for rapid discrimination of early-stage regulated cell death modalities

Real-time nanomechanical visualization and profiling for rapid discrimination of early-stage regulated cell death modalities

Pan, Shaoshan; Jia, Haiying; Fang, Xue; Liu, Qian; Wang, Qian; Luo, Tianzhi

DOI: PMID:

Abstract

Accurate and early discrimination of regulated cell death (RCD) modalities, such as ferroptosis, apoptosis, and cuproptosis, remains a critical challenge for advancing disease understanding and therapy. Existing analytical approaches often lack the capabilities for real-time, non-invasive, and discriminative monitoring. Here, we introduce TFM-CRIM, an innovative nanomechanical profiling platform. It synergistically integrates traction force microscopy (TFM) for visualizing and profiling cellular nanomechanics with confocal reflection interference microscopy (CRIM), ensuring precise cell boundary definition for enhanced accuracy in force quantification. This platform enables label-free, quantitative, and dynamic assessment of nanomechanical responses in early RCD. Real-time TFM-CRIM monitoring revealed unique kinetic signatures discriminating RCD modalities: staurosporine-induced apoptosis exhibited an extremely rapid force collapse to near-zero by ∼20–30 min (initiating within 10 min); Elesclomol-CuCl2-induced cuproptosis displayed a swift, sustained force decrease from ∼10 to 20 min, stabilizing at a low level by ∼40–60 min. These profiles were markedly different from the slower ferroptosis kinetics: RSL3-induction initiated a sustained force reduction ∼20–40 min (over ∼3 h), while Erastin-induction, after an early subtle decrease from ∼20 min, showed a delayed major decline from ∼120 min (spanning ∼6 h). Critically, TFM-CRIM demonstrated significantly superior sensitivity and temporal resolution compared to nanoindentation (delayed/non-continuous) and cellular impedance measurements (slower, less distinct kinetics). These distinct nanomechanical signatures, correlated with alterations in cytoskeletal actin fibres, enable robust RCD classification. This work establishes TFM-CRIM-based nanomechanical profiling as a rapid, quantitative, and highly sensitive analytical method for early RCD characterization, poised to accelerate mechanistic insights and drug screening.

Keywords

Nanomechanical profiling ; Regulated cell death ; Cell traction force ; Label-free analysis ; Dynamic response ; Actin cytoskeleton

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