The mechanical properties of tissues and cells have proven to be of great importance in biology and medicine, spawning major research efforts and commercial outcomes. The interplay between mechanical and biochemical cues and the mechanical properties of subcellular and cellular constituents has come to be appreciated as key to understanding many fundamental aspects of biology as well as the genesis and progression of disease. At the same time, at the other extreme, the mechanical properties of whole organs and their constituents have been shown to be effective markers of disease on the scale of the human body, as probed by ultrasound elastography and magnetic resonance elastography, both of which, after 20-year gestations, have reached commercial markets.

Amyloidopathy is one of the most prominent hallmarks of Alzheimer's disease (AD), the leading cause of dementia worldwide, and is characterized by the accumulation of amyloid plaques in the brain parenchyma. The plaques consist of abnormal deposits mainly composed of an aggregationprone protein fragment, β-amyloid 1-40/1-42, into the extracellular matrix. Brillouin microspectroscopy is an all-optical contactless technique that is based on the interaction between visible light and longitudinal acoustic waves or phonons, giving access to the viscoelasticity of a sample on a subcellular scale. Here, we describe the first application of micromechanical mapping based on Brillouin scattering spectroscopy to probe the stiffness of individual amyloid plaques in the hippocampal part of the brain of a β-amyloid overexpressing transgenic mouse. Correlative analysis based on Brillouin and Raman microspectroscopy showed that amyloid plaques have a complex structure with a rigid core of β-pleated sheet conformation (β-amyloid) protein surrounded by a softer ring-shaped region richer in lipids and other protein conformations. These preliminary results give a new insight into the plaque biophysics and biomechanics, and a valuable contrast mechanism for the study and diagnosis of amyloidopathy.

Brillouin imaging (BI) for micromechanical characterization of tissues and biomaterials is a fast-developing field of research with a strong potential for medical diagnosis of disease-modified tissues and cells. Although the principles of BI imply its compatibility with in vivo and in situ measurements, the integration of BI with a flexible catheter, capable of reaching the region of interest within the body, is yet to be reported. Here, for the first time, we experimentally investigate integration of the Brillouin spectroscope with standard optical fiber components to achieve a Brillouin endoscope. The performance of single-fiber and dual-fiber endoscopes are demonstrated and analyzed. We show that a major challenge in construction of Brillouin endoscopes is the strong backward Brillouin scattering in the optical fiber and we present a dual-fiber geometry as a possible solution. Measurements of Brillouin spectra in test liquids (water, ethanol and glycerol) are demonstrated using the dual-fiber endoscope and its performance is analyzed numerically with the help of a beam propagation model.

Tumor growth is regulated by a diverse set of extracellular influences, including paracrine crosstalk with stromal partners, and biophysical interactions with surrounding cells and tissues. Studies elucidating the role of physical force and the mechanical properties of the extracellular matrix (ECM) itself as regulators of tumor growth and invasion have been greatly catalyzed by the use of in vitro three-dimensional (3D) tumor models. These systems provide the ability to systematically isolate, manipulate, and evaluate impact of stromal components and extracellular mechanics in a platform that is both conducive to imaging and biologically relevant. However, recognizing that mechanoregulatory crosstalk is bi-directional and fully utilizing these models requires complementary methods for in situ measurements of the local mechanical environment. Here, in 3D tumor/fibroblast co-culture models of pancreatic cancer, a disease characterized by its prominent stromal involvement, we evaluate the use of particle-tracking microrheology to probe dynamic mechanical changes. Using videos of fluorescently labeled polystyrene microspheres embedded in collagen I ECM, we measure spatiotemporal changes in the Brownian motion of probes to report local ECM shear modulus and microheterogeneity. This approach reveals stiffening of collagen in fibroblast co-cultures relative to cultures with cancer cells only, which exhibit degraded ECM with heterogeneous microstructure. We further show that these effects are dependent on culture geometry with contrasting behavior for embedded and overlay cultures. In addition to potential application to screening stroma-targeted therapeutics, this work also provides insight into how the composition and plating geometry impact the mechanical properties of 3D cell cultures that are increasingly widely used in cancer biology.

Brillouin spectroscopy is a powerful tool for measuring the mechanical properties of materials without contact. The sensitivity to mechanical changes that a Brillouin spectrometer can detect is determined by the precision to which a spectral peak can be localized. The localization precision is however fundamentally limited by the low number of photons within a Brillouin measurement, as well as by intrinsic noise of the setup. Here, we present a method to improve the spectral sensitivity of Brillouin measurements by exploiting the autocorrelation function of the spectrum. We show that by performing a localization process on the autocorrelation function nearly 20% increase in localization precision can be obtained. This result is consistent between our theoretical treatment, numerical simulation and experimental results. We further study the effect of background noise on the precision improvement for realistic scenarios.

The three-dimensional (3D) mechanical response of the cornea to intraocular pressure (IOP) elevation has not been previously reported. In this study, we use an ultrasound speckle tracking technique to measure the 3D displacements and strains within the central 5.5mm of porcine corneas during the whole globe inflation. Inflation tests were performed on dextran-treated corneas (treated with a 10% dextran solution) and untreated corneas. The dextran-treated corneas showed an inflation response expected of a thin spherical shell, with through-thickness thinning and in-plane stretch, although the strain magnitudes exhibited a heterogeneous spatial distribution from the central to more peripheral cornea. The untreated eyes demonstrated a response consistent with swelling during experimentation, with through-thickness expansion overriding the inflation response. The average volume ratios obtained in both groups was near 1 confirming general incompressibility, but local regions of volume loss or expansion were observed. These results suggest that biomechanical measurements in 3D provide important new insight to understand the mechanical response of ocular tissues such as the cornea.

In this paper, we point out some practical obstacles arising in realization of compressional optical coherence elastography (OCE) that have not attracted su±cient attention previously. Specifically, we discuss (i) complications in quantification of the Young modulus of tissues related to partial adhesion between the OCE probe and soft intervening reference layer sensor, (ii) distorting influence of tissue surface curvature/corrugation on the subsurface strain distribution mapping, (iii) ways of signal-to-noise ratio (SNR) enhancement in OCE strain mapping when periodic averaging is not realized, and (iv) potentially significant influence of tissue elastic nonlinearity on quantification of its stiffness. Potential practical approaches to mitigate the effects of these complications are also described.

We present a multimodal ferrule-top sensor designed to perform the integrated epidetection of Optical Coherence Tomography (OCT) depth-profiles and micron-scale indentation by all-optical detection. By scanning a sample under the probe, we can obtain structural cross-section images and identify a region-of-interest in a nonhomogeneous sample. Then, with the same probe and setup, we can immediately target that area with a series of spherical-indentation measurements, in which the applied load is known with a μN precision, the indentation depth with sub-μm precision and a maximum contact radius of 100 μm. Thanks to the visualization of the internal structure of the sample, we can gain a better insight into the observed mechanical behavior. The ability to impart a small, confined load, and perform OCT A-scans at the same time, could lead to an alternative, high transverse resolution, Optical Coherence Elastography (OCE) sensor.

In the characterization of elastic properties of tissue using dynamic optical coherence elastography, shear/surface waves are propagated and tracked in order to estimate speed and Young's modulus. However, for dispersive tissues, the displacement pulse is highly damped and distorted during propagation, diminishing the effectiveness of peak tracking approaches, and leading to biased estimates of wave speed. Further, plane wave propagation is sometimes assumed, which contributes to estimation errors. Therefore, we invert a wave propagation model that incorporates propagation, decay, and distortion of pulses in a dispersive media in order to accurately estimate its elastic and viscous components. The model uses a general first-order approximation of dispersion, avoiding the use of any particular rheological model of tissue. Experiments are conducted in elastic and viscoelastic tissue-mimicking phantoms by producing a Gaussian push using acoustic radiation force excitation and measuring the wave propagation using a Fourier domain optical coherence tomography system. Results confirmed the effectiveness of the inversion method in estimating viscoelastic parameters in both the viscoelastic and elastic phantoms when compared to mechanical measurements. Finally, the viscoelastic characterization of a fresh porcine cornea was conducted. Preliminary results validate this approach when compared to other methods.

The ocular lens stifiens dramatically with age, resulting in a loss of function. However, the mechanism of stifiening remains unknown, at least in part due to di±culties in making reliable measurements of the intrinsic mechanical properties of the lens. Recent experiments have employed manual compression testing to evaluate the stifiness of murine lenses which have genotypes pertinent to human lens diseases. These experiments compare the extrinsic stifiness of lenses from the genotype of interest to the wild-type lens in an efiort to reach conclusions regarding the cellular or molecular basis of lens stifiening. However, these comparisons are confounded by alterations in lens size and geometry which invariably accompany these genetic manipulations. Here, we utilize manual lens compression to characterize the stifiness of a porcine lens and a murine lens. An inverse elastographic technique was then developed to estimate the intrinsic shear modulus of each lens as well as the elastic modulus of the lens capsule. The results were in good agreement with the previous literature values.

On page 1650021-7, in Appendix B, in the first line “Ref. 23" should be “Ref. 13".