Evaluation of commercial virtually imaged phase array and Fabry-Pérot based Brillouin spectrometers for applications...

Measuring the complex mechanical properties of biological objects has become a necessity to answer key questions in mechanobiology and to propose innovative clinical and therapeutic strategies. In this context, Brillouin light scattering (BLS) has recently come into vogue, offering quantitative imaging of the mechanical properties without labels and with a micrometer resolution. In biological samples, the magnitude of the spectral changes are typically of a few tens of MHz, and the ability of modern spectrometers to monitor such subtle changes needs to be evaluated. Moreover, the multiplicity of variations in optical arrangements, specific to each lab, requires to set a standard for the assessment of the characteristics of BLS systems. In this paper we propose a protocol to evaluate the precision and accuracy of two commercial spectrometers that is reproducible across labs. For a meaningful comparison, we coupled the spectrometers to the same microscope and to the same laser. We first evaluated the optimum acquisition time and laser power. We evaluated the precision using pure water samples. We determined the accuracy by probing water solutions with increasing concentration of salt and comparing it with theory. Following these quantifications, we applied the VIPA-based spectrometer to tumor spheroids engineered from different cell lines that possess different metastatic potentials and resistance to therapies. On these models, we detected significant changes in the linewidth suggesting that BLS measurements of the viscosity could be used as a read-out to distinguish different levels of drug resistance.