Separating water content from network dynamics in cell nuclei with Brillouin microscopy

Abstract

Probing forces, deformations and generally speaking the mechanical properties of cells is the hallmark of mechanobiology. In the last two decades many techniques have been developed to this end that are largely based on deforming the cells and measuring the reaction force. In cells, an alternative approach has been implemented mid 2010's, based on Brillouin Light Scattering (BLS) that produces a spectrum that can be interpreted as the response of the sample to an infinitesimal uniaxial compression at picosecond timescales. In all of these measurements, the response of the cell is quantified with a colloquial "stiffness" that encompasses both the contribution of load-bearing structures and volume changes, much to confusion. To clarify the interpretation of the hypersonic data obtained from BLS spectra, we vary the relative volume fraction of intracellular water and solid network by applying osmotic compressions to single cells. In the nucleus, we observe a non-linear increase in the sound velocity and attenuation with increasing osmotic pressure that we fit to a poroelastic model, providing an estimate of the friction coefficient between the water phase and the network. By comparing BLS data to volume measurements, our approach demonstrates clearly that BLS shift alone is mostly sensitive to water content while the additional analysis of the linewidth allows identifying the contribution of the biopolymer-based network dynamics in living cells.