Gelatin hydrogels cover a wide range of microenvironments that are relevant for bioapplications of Brillouin elastography.

Brillouin and Raman scattering spectroscopy are established techniques for the nondestructive contactless and label-free readout of mechanical, chemical, and structural properties of condensed matter. Brillouin-Raman investigations currently require separate measurements and a site-matched approach to obtain complementary information from a sample. Here, we demonstrate a new concept of fully scanning multimodal microspectroscopy for simultaneous detection of Brillouin and Raman light scattering in an exceptionally wide spectral range, from fractions of GHz to hundreds of THz. It yields an unprecedented 150-dB contrast, which is especially important for the analysis of opaque or turbid media such as biomedical samples, and spatial resolution on a subcellular scale. We report the first applications of this new multimodal method to a range of systems, from a single cell to the fast reaction kinetics of a curing process, and the mechanochemical mapping of highly scattering biological samples.

When a solute is dissolved in water, their mutual interactions determine the molecular properties of the solute on one hand, and the structure and dynamics of the surrounding water particles (the so-called hydration water) on the other. The very existence of soft matter and its peculiar properties are largely due to the wide variety of possible water–solute interactions. In this context, water is not an inert medium but rather an active component, and hydration water plays a crucial role in determining the structure, stability, dynamics, and function of matter. This review focuses on the collective dynamics of hydration water in terms of retardation with respect to the bulk, and of the number of molecules whose dynamics is perturbed. Since water environments are in a dynamic equilibrium, with molecules continuously exchanging from around the solute towards the bulk and vice versa, we examine the ability of different techniques to measure the water dynamics on the basis of the explored time scales and exchange rates. Special emphasis is given to the collective dynamics probed by extended depolarized light scattering and we discuss whether and to what extent the results obtained in aqueous solutions of small molecules can be extrapolated to the case of large biomacromolecules. In fact, recent experiments performed on solutions of increasing complexity clearly indicate that a reductionist approach is not adequate to describe their collective dynamics. We conclude this review by presenting current ideas that are being developed to describe the dynamics of water interacting with macromolecules.

In every biological tissue, morphological and topological properties strongly affect its mechanical features and behaviour, so that ultrastructure, composition and mechanical parameters are intimately connected. Overall, it is their correct interplay that guarantees the tissue functionality. The development of experimental methods able to correlate these properties would open new opportunities both in the biological and the biomedical fields. Here, we report a correlative study intended to map supramolecular morphology, biochemical composition and viscoelastic parameters of collagen by all-optical microscopies. In particular, using human corneal tissue as a benchmark, we correlate Second-Harmonic Generation maps with mechanical and biochemical imaging obtained by Brillouin and Raman micro-spectroscopy. The study highlights how subtle variations in supramolecular organization originate the peculiar mechanical behavior of different subtypes of corneal lamellae. The presented methodology paves the way to the non-invasive assessment of tissue morpho-mechanics in biological as well as synthetic materials.

Brillouin imaging is an emerging optical elastography technique that is able to generate maps of the mechanical properties at microscale with great potential in biophysical and biomedical fields. A key parameter is its spatial resolution, which is usually identified with that of the confocal microscope coupled to the Brillouin interferometer. Conversely, here we demonstrate that the mean free path of acoustic phonons plays a major role in defining the resolution, especially for high numerical aperture confocal setups. Surprisingly, the resolution of elastography maps may even deteriorate when decreasing the scattering volume.

Brillouin light scattering (BLS) spectroscopy is a non-invasive, non-contact, label-free optical technique that can provide information on the mechanical properties of a material on the submicrometre scale. Over the past decade, BLS has found increasing microscopy applications in the life sciences, driven by the observed importance of mechanical properties in biological processes, the realization of more sensitive BLS spectrometers and the extension of BLS to an imaging modality. As with other spectroscopic techniques, BLS measurements detect not only signals that are characteristic of the investigated sample, but also those of the experimental apparatus, and can be substantially affected by measurement conditions. Here we report a consensus between researchers in the field. We aim to improve the comparability of BLS studies by providing reporting recommendations for the measured parameters and detailing common artefacts. Given that most BLS studies of biological matter are still at proof-of-concept stages and use different, often self-built, spectrometers, a consensus statement is particularly timely to ensure unified advancement.