Correlation of biomechanics and cancer cell phenotype by combined Brillouin and Raman spectroscopy of U87-MG glioblastoma cells.

The elucidation of biomechanics furthers our understanding of brain tumour biology. Brillouin spectroscopy is a new optical method that addresses viscoelastic properties down to subcellular resolution in a contact-free manner. Moreover, it can be combined with Raman spectroscopy to obtain co-localized biochemical information. Here, we applied co-registered Brillouin and Raman spectroscopy to U87-MG human glioblastoma cells in vitro. Using two-dimensional and three-dimensional cultures, we related biomechanical properties to local biochemical composition at the subcellular level, as well as the cell phenotype. Brillouin and Raman mapping of adherent cells showed that the nucleus and nucleoli are stiffer than the perinuclear region and the cytoplasm. The biomechanics of the cell cytoplasm is affected by culturing conditions, i.e. cells grown as spheroids are stiffer than adherent cells. Inside the spheroids, the presence of lipid droplets as assessed by Raman spectroscopy revealed higher Brillouin shifts that are not related to a local increase in stiffness, but are due to a higher refractive index combined with a lower mass density. This highlights the importance of locally defined biochemical reference data for a correct interpretation of the Brillouin shift of cells and tissues in future studies investigating the biomechanics of brain tumour models by Brillouin spectroscopy.

Brillouin and Raman imaging of domain walls in periodically-poled 5%-MgO:LiNbO3.

Recently, ferroelectric domain walls (DWs) have attracted considerable attention due to their intrinsic topological effects and their huge potential for optoelectronic applications. In contrast, many of the underlying physical properties and phenomena are not well characterized. In this regard, analyzing the vibrational properties, e.g. by Raman spectroscopy, provides direct access to the various local material properties, such as strains, defects or electric fields. While the optical phonon spectra of DWs have been widely investigated in the past, no reports on the acoustic phonon properties of DWs exist. In this work, we present a joint Raman and Brillouin visualization of ferroelectric DWs in the model ferroelectric lithium niobate. This is possible by using a combined Raman and virtually imaged phased array Brillouin setup. Here, we show that DWs can be visualized via frequency shifts observed in the acoustic phonons, as well. The observed contrast then is qualitatively explained by models adapted from Raman spectroscopy. This work, hence, provides a novel route to study ferroelectric DWs and their intrinsic mechanical properties.

Quantifying the refractive index of ferroelectric domain walls in periodically poled LiNbO3 single crystals by polarization-sensitive optical coherence tomography.

Domain walls (DWs) in ferroelectric (FE) and multiferroic materials possess an ever-growing potential as integrated functional elements, for instance in optoelectronic nanodevices. Mandatory, however, is the profound knowledge of the local-scale electronic and optical properties, especially at DWs that are still incompletely characterized to date. Here, we quantify the refractive index of individual FE DWs in periodically-poled LiNbO3 (PPLN) single crystals. When applying polarization-sensitive optical coherence tomography (PS-OCT) at 1300 nm using circular light polarization, we are able to probe the relevant electro-optical properties close to and at the DWs, including also their ordinary and extraordinary contributions. When comparing to numerical calculations, we conclude that the DW signals recorded for ordinary and extraordinary polarization stem from an increased refractive index of at least Δn > 2·10-3 that originates from a tiny region of < 30 nm in width. PS-OCT hence provides an extremely valuable tool to decipher and quantify subtle changes of refractive index profiles for both inorganic and biomedical nanomaterial systems.

Brillouin confocal microscopy to determine biomechanical properties of SULEEI-treated bovine pericardium for application in cardiac surgery.

BACKGROUND: Heart valves are exposed to a highly dynamic environment and underlie high tensile and shear forces during opening and closing. Therefore, analysis of mechanical performance of novel heart valve bioprostheses materials, like SULEEI-treated bovine pericardium, is essential and usually carried out by uniaxial tensile tests. Nevertheless, major drawbacks are the unidirectional strain, which does not reflect the in vivo condition and the deformation of the sample material. An alternative approach for measurement of biomechanical properties is offered by Brillouin confocal microscopy (BCM), a novel, non-invasive and three-dimensional method based on the interaction of light with acoustic waves. OBJECTIVE: BCM is a powerful tool to determine viscoelastic tissue properties and is, for the first time, applied to characterize novel biological graft materials, such as SULEEI-treated bovine pericardium. Therefore, the method has to be validated as a non-invasive alternative to conventional uniaxial tensile tests. METHODS: Vibratome sections of SULEEI-treated bovine pericardium (decellularized, riboflavin/UV-cross-linked and low-energy electron irradiated) as well as native and GA-fixed controls (n = 3) were analyzed by BCM. In addition, uniaxial tensile tests were performed on equivalent tissue samples and Young's modulus as well as length of toe region were analyzed from stress-strain diagrams. The structure of the extracellular matrix (ECM), especially collagen and elastin, was investigated by multiphoton microscopy (MPM). RESULTS: SULEEI-treated pericardium exhibited a significantly higher Brillouin shift and hence higher tissue stiffness in comparison to native and GA-fixed controls (native: 5.6±0.2 GHz; GA: 5.5±0.1 GHz; SULEEI: 6.3±0.1 GHz; n = 3, p < 0.0001). Similarly, a significantly higher Young's modulus was detected in SULEEI-treated pericardia in comparison to native tissue (native: 30.0±10.4 MPa; GA: 31.8±10.7 MPa; SULEEI: 42.1±7.0 MPa; n = 3, p = 0.027). Native pericardia showed wavy and non-directional collagen fibers as well as thin, linear elastin fibers generating a loose matrix. The fibers of GA-fixed and SULEEI-treated pericardium were aligned in one direction, whereat the SULEEI-sample exhibited a much denser matrix. CONCLUSION: BCM is an innovative and non-invasive method to analyze elastic properties of novel pericardial graft materials with special mechanical requirements, like heart valve bioprostheses.

Ferroelastic domain identification in BiFeO3 crystals using Raman spectroscopy.

Multiferroic BiFeO3 crystals were investigated by means of micro-Raman spectroscopy using the laser wavelengths of 442 nm (resonant conditions) and 633 nm (non-resonant conditions). The azimuthal angle dependence of the intensity of the Raman modes allowed their symmetry assignment. The experimental data are consistent with a simulation based on Raman tensor formalism. Mixed symmetries were taken into account, considering the orientation of the crystal optic axis along a pseudo-cubic <111> direction. The strong anisotropic intensity variation of some of the polar Raman modes was used for line scans and mappings in order to identify ferroelastic domain patterns. The line scans performed with different excitation wavelengths and hence different information depths indicate a tilt of the domain walls with respect to the sample surface. The domain distribution found by Raman spectroscopy is in very good agreement with the finding of electron back scattering diffraction.