RESUMO
Lamin A/C is a well-established key contributor to nuclear stiffness and its role in nucleus mechanical properties has been extensively studied. However, its impact on whole-cell mechanics has been poorly addressed, particularly concerning measurable physical parameters. In this study, we combined microfluidic experiments with theoretical analyses to quantitatively estimate the whole-cell mechanical properties. This allowed us to characterize the mechanical changes induced in cells by lamin A/C alterations and prelamin A accumulation resulting from atazanavir treatment or lipodystrophy-associated LMNA R482W pathogenic variant. Our results reveal a distinctive increase in long-time viscosity as a signature of cells affected by lamin A/C alterations. Furthermore, they show that the whole-cell response to mechanical stress is driven not only by the nucleus but also by the nucleo-cytoskeleton links and the microtubule network. The enhanced cell viscosity assessed with our microfluidic assay could serve as a valuable diagnosis marker for lamin-related diseases.
RESUMO
In this article, we show an alternative low-cost fabrication method to obtain poly(dimethyl siloxane) (PDMS) microfluidic devices. The proposed method allows the inscription of micron resolution channels on polystyrene (PS) surfaces, used as a mold for the wanted microchip's production, by applying a high absorption coating film on the PS surface to ablate it with a focused low-power visible laser. The method allows for obtaining micro-resolution channels at powers between 2 and 10 mW and can realize any two-dimensional polymeric devices. The effect of the main processing parameters on the channel's geometry is presented.