Modeling stem cell nucleus mechanics using confocal microscopy.

Nuclear mechanics is emerging as a key component of stem cell function and differentiation. While changes in nuclear structure can be visually imaged with confocal microscopy, mechanical characterization of the nucleus and its sub-cellular components require specialized testing equipment. A computational model permitting cell-specific mechanical information directly from confocal and atomic force microscopy of cell nuclei would be of great value. Here, we developed a computational framework for generating finite element models of isolated cell nuclei from multiple confocal microscopy scans and simple atomic force microscopy (AFM) tests. Confocal imaging stacks of isolated mesenchymal stem cells were converted into finite element models and siRNA-mediated Lamin A/C depletion isolated chromatin and Lamin A/C structures. Using AFM-measured experimental stiffness values, a set of conversion factors were determined for both chromatin and Lamin A/C to map the voxel intensity of the original images to the element stiffness, allowing the prediction of nuclear stiffness in an additional set of other nuclei. The developed computational framework will identify the contribution of a multitude of sub-nuclear structures and predict global nuclear stiffness of multiple nuclei based on simple nuclear isolation protocols, confocal images and AFM tests.

Fabrication and Performance of a Multi-Discharge Cold Atmospheric Pressure Plasma Array.

Cold atmospheric pressure plasma (CAP) has been shown to kill bacteria and remove biofilms. Here we report the development of a unique CAP array device consisting of a parallel stack of eight linear-discharge plasma elements that create a ~ 5 cm2 (2.4 cm × 2 cm) treatment area. The CAP device is fabricated from Low Temperature Co-fired Ceramic (LTCC) layers to create 24 mm long linear-discharge channels (500 µm gap) with embedded opposing silver metal electrodes. A 20 kHz AC voltage (0.5-5 kV) applied to the electrodes generates an Ar/O2 plasma between the plates, with the gas flow directing the reactive species toward the biological sample (biofilms, etc.) to affect the antimicrobial treatment. External ballast resistors were used to study discharge uniformity in the stacked array elements and internal thick film ballast resistors (≈150 kΩ) were developed to create a fully integrated device. Typical element discharge currents were 1-2.5 mA with the total array current tested at 20 mA to provide optimal device uniformity. The plasma discharge was further shown to produce reactive hydrogen peroxide and exert antimicrobial effects on Pseudomonas biofilms and Salmonella contaminated eggshell samples, with >99% of the bacterial cells killed with less than 60 seconds of plasma exposure.