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High-resolution assessment of multidimensional cellular mechanics using label-free refractive-index traction force microscopy
oleh: Moosung Lee, Hyuntae Jeong, Chaeyeon Lee, Mahn Jae Lee, Benedict Reve Delmo, Won Do Heo, Jennifer H. Shin, YongKeun Park
Format: | Article |
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Diterbitkan: | Nature Portfolio 2024-01-01 |
Deskripsi
Abstract A critical requirement for studying cell mechanics is three-dimensional assessment of cellular shapes and forces with high spatiotemporal resolution. Traction force microscopy with fluorescence imaging enables the measurement of cellular forces, but it is limited by photobleaching and a slow acquisition speed. Here, we present refractive-index traction force microscopy (RI-TFM), which simultaneously quantifies the volumetric morphology and traction force of cells using a high-speed illumination scheme with 0.5-Hz temporal resolution. Without labelling, our method enables quantitative analyses of dry-mass distributions and shear (in-plane) and normal (out-of-plane) tractions of single cells on the extracellular matrix. When combined with a constrained total variation-based deconvolution algorithm, it provides 0.55-Pa shear and 1.59-Pa normal traction sensitivity for a 1-kPa hydrogel substrate. We demonstrate its utility by assessing the effects of compromised intracellular stress and capturing the rapid dynamics of cellular junction formation in the spatiotemporal changes in non-planar traction components.