Label-Free Optical Mapping for Large-Area Biomechanical Dynamics of Multicellular Systems.

Mapping cellular activities over large areas is crucial for understanding the collective behaviors of multicellular systems. Biomechanical properties, such as cellular traction force, serve as critical regulators of physiological states and molecular configurations. However, existing technologies for mapping large-area biomechanical dynamics are limited by the small field of view and scanning nature. To address this, we propose a novel platform that utilizes a vast number of optical diffractive elements for mapping large-area biomechanical dynamics. This platform achieves a field-of-view of 10.6 mm X 10.6 mm, a three-orders-of-magnitude improvement over traditional traction force microscopy. Transient mechanical waves generated by monolayer neonatal rat ventricular myocytes were captured with high spatiotemporal resolution (130 fps and 20 µm for temporal and spatial resolution, respectively). Furthermore, its label-free nature allows for long-term observations extended to a week, with minimal disruption of cellular functions. Finally, simultaneous measurements of calcium ions concentrations and biomechanical dynamics are demonstrated.

Simulation of GHz ultrasonic wave piezoelectric instrumentation for Fourier transform computation.

The recent emerging alternative to classic numerical Fast Fourier transform (FFT) computation, based on GHz ultrasonic waves generated from and detected by piezoelectric transducers for wavefront computing (WFC), is more efficient and energy-saving. In this paper, we present comprehensive studies on the modeling and simulation methods for ultrasonic WFC computation. We validate the design of the WFC system using ray-tracing, Fresnel diffraction (FD), and the full-wave finite element method (FEM). To effectively simulate the WFC system for inputs of 1-D signals and 2-D images, we verified the design parameters and focal length of an ideal plano-concave lens using the ray-tracing method. We also compared the analytical FFT solution with our Fourier transform (FT) results from 3-D and 2-D FD and novel 2-D full wave FEM simulations of a multi-level Fresnel lens with 1-D signals and 2-D images as inputs. Unlike the previously reported WFC system which catered only for 2-D images, our proposed method also can solve the 1-D FFT effectively. We validate our proposed 2-D full wave FEM simulation method by comparing our results with the theoretical FFT and Fresnel diffraction method. The FFT results from FD and FEM agree well with the digitally computed FFT, with computational complexity reduced from [Formula: see text] to O(N) for 2-D FFT, and from O(NlogN) to O(N) for 1-D FFT with a large number of signal sampling points N.

Distributed colorimetric interferometer for mapping the pressure distribution in a complex microfluidics network.

We demonstrate a novel platform for mapping the pressure distribution of complex microfluidics networks with high spatial resolution. Our approach utilizes colorimetric interferometers enabled by lossy optical resonant cavities embedded in a silicon substrate. Detection of local pressures in real-time within a fluid network occurs by monitoring a reflected color emanating from each optical cavity. Pressure distribution measurements spanning a 1 cm2 area with a spatial resolution of 50 µm have been achieved. We applied a machine-learning-assisted sensor calibration method to generate a dynamic measurement range from 0 to 5.0 psi, with 0.2 psi accuracy. Adjustments to this dynamic measurement range are possible to meet different application needs for monitoring flow conditions in complex microfluidics networks, for the timely detection of anomalies such as clogging or leakage at their occurring locations.

Type V Collagen in Scar Tissue Regulates the Size of Scar after Heart Injury.

Scar tissue size following myocardial infarction is an independent predictor of cardiovascular outcomes, yet little is known about factors regulating scar size. We demonstrate that collagen V, a minor constituent of heart scars, regulates the size of heart scars after ischemic injury. Depletion of collagen V led to a paradoxical increase in post-infarction scar size with worsening of heart function. A systems genetics approach across 100 in-bred strains of mice demonstrated that collagen V is a critical driver of postinjury heart function. We show that collagen V deficiency alters the mechanical properties of scar tissue, and altered reciprocal feedback between matrix and cells induces expression of mechanosensitive integrins that drive fibroblast activation and increase scar size. Cilengitide, an inhibitor of specific integrins, rescues the phenotype of increased post-injury scarring in collagen-V-deficient mice. These observations demonstrate that collagen V regulates scar size in an integrin-dependent manner.

Differential Contributions of Actin and Myosin to the Physical Phenotypes and Invasion of Pancreatic Cancer Cells.

INTRODUCTION: Metastasis is a fundamentally physical process in which cells deform through narrow gaps and generate forces to invade surrounding tissues. While it is commonly thought that increased cell deformability is an advantage for invading cells, we previously found that more invasive pancreatic ductal adenocarcinoma (PDAC) cells are stiffer than less invasive PDAC cells. Here we investigate potential mechanisms of the simultaneous increase in PDAC cell stiffness and invasion, focusing on the contributions of myosin II, Arp2/3, and formins. METHOD: We measure cell invasion using a 3D scratch wound invasion assay and cell stiffness using atomic force microscopy (AFM). To determine the effects of actin- and myosin-mediated force generation on cell stiffness and invasion, we treat cells with pharmacologic inhibitors of myosin II (blebbistatin), Arp2/3 (CK-666), and formins (SMIFH2). RESULTS: We find that the activity of myosin II, Arp2/3, and formins all contribute to the stiffness of PDAC cells. Interestingly, we find that the invasion of PDAC cell lines is differentially affected when the activity of myosin II, Arp2/3, or formins is inhibited, suggesting that despite having similar tissue origins, different PDAC cell lines may rely on different mechanisms for invasion. CONCLUSIONS: These findings deepen our knowledge of the factors that regulate cancer cell mechanotype and invasion, and incite further studies to develop therapeutics that target multiple mechanisms of invasion for improved clinical benefit.

Plasmonic micropillars for precision cell force measurement across a large field-of-view.

A plasmonic micropillar platform with self-organized gold nanospheres is reported for the precision cell traction force measurement across a large field-of-view (FOV). Gold nanospheres were implanted into the tips of polymer micropillars by annealing gold microdisks with nanosecond laser pulses. Each gold nanosphere is physically anchored in the center of a pillar tip and serves as a strong, point-source-like light scattering center for each micropillar. This allows a micropillar to be clearly observed and precisely tracked even under a low magnification objective lens for the concurrent and precision measurement across a large FOV. A spatial resolution of 30 nm for the pillar deflection measurement has been accomplished on this platform with a 20× objective lens.