This PDF file contains the front matter associated with SPIE Proceedings Volume 9710, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

Optical coherence elastography (OCE) is emerging as a potentially useful tool in the identification of a number of diseases. In our group, we are developing OCE techniques based on compressive loading. Typically, these techniques employ a quasi-static mechanical load introduced by uniaxially compressing a sample with a rigid plate. The resulting deformation of the sample is measured using phase-sensitive detection and the local axial strain is estimated from the slope of displacement over a finite depth in the sample, providing qualitative mechanical contrast. In this talk, an overview of our work will be given and some of the outstanding challenges described. Our group’s work in OCE can broadly be divided into four streams, each of which will be described in detail in the talk: system development; techniques; quantification; and applications. • System development: The phase-sensitive OCE method we have developed will be described, as well as a high resolution optical coherence microscopy-based elastography system suitable for imaging cellular-scale mechanical properties. • Techniques: In addition to presenting techniques to estimate strain, our approaches to imaging tissue viscoelasticity and nonlinearity will be described. A technique to segment elastograms based on strain heterogeneity will be presented. • Quantification: Methods under development to quantify tissue stiffness in compression OCE will be described. This work is enabled by optical palpation and solutions to the forward and inverse elasticity problems. • Applications: Three applications areas will be described: intraoperative assessment of tumour margins, mapping stiffness in tumour biology and assessing the stiffness of cardiovascular tissue in an animal model.

The stiffness or rigidity of the extracellular matrix (ECM) regulates cell response. Established mechanical tests to measure stiffness, such as indentation and tensile tests, are invasive and destructive to the sample. Endogenous or native molecules to cells and ECM components, like tryptophan and cross-links of collagen, display fluorescence upon irradiation with ultraviolet light. Most likely, the concentration of these endogenous fluorophores changes as the stiffness of the ECM changes. In this work we investigate the endogenous fluorescence of collagen gels containing fibroblasts as a non-invasive non-destructive method to measure stiffness of the ECM. Human fibroblast cells were cultured in three-dimensional gels of type I collagen (50,000 cells/ml). This construct is a simple model of tissue contraction. During contraction, changes in the excitation-emission matrix (a fluorescence map in the 240-520/290-530 nm range) of constructs were measured with a spectrofluoremeter, and changes in stiffness were measured with a standard indentation test over 16 days. Results show that a progressive increase in fluorescence of the 290/340 nm excitation-emission pair correlates with a progressive increase in stiffness (r=0.9, α=0.5). The fluorescence of this excitation-emission pair is ascribed to tryptophan and variations in the fluorescence of this pair correlate with cellular proliferation. In this tissue model, the endogenous functional fluorescence of proliferating fibroblast cells is a biomechanical marker of stiffness of the ECM.

Mechanical preconditioning and mechanical properties of tissue engineered constructs are essential for their capability to regenerate damaged tissues. To online monitor the mechanical properties a hydrostatic pressure bioreactor was coupled with optical coherence tomography into a new image modality termed hydrostatic pressure optical coherence elastography (HP-OCE). HP-OCE was utilised to assess the properties of three-dimensional (3D) tissue constructs while being physically stimulated within the hydrostatic force bioreactor. Hydrogels have been infiltrated into porous rapid prototyped or salt-leached scaffolds to mimic heterogeneous mechanical properties of cell-seeded constructs. Variations of mechanical properties in the solid scaffolds and agarose gels with different gel concentrations as well as the presences of cells have been clearly delineated by HP-OCE. Results indicate that HP-OCE allows contactless real-time non-invasive monitoring of the mechanical properties of tissue constructs and the effect of physical stimulation on cellular activities.

The stiffness of the extra cellular matrix (ECM) is recognized as a key regulator of cancer cell proliferation, migration and invasion. Therefore technologies that quantify ECM stiffness with micro-scale scale resolution will likely provide important insights into neoplastic progression. Laser Speckle Micro-Rheology (LSM) is a novel optical tool for measuring tissue viscoelastic properties with micro-scale resolution. In LSM, speckle images are collected through an objective lens by a high-speed camera. Spatio-temporal correlation analysis of speckle frames yields the intensity autocorrelation function, g2(t), for each pixel, and subsequently a 2D map of viscoelastic modulus, G*(ω) is reconstructed. Here, we investigate the utility of LSM for micro-mechanical evaluation of the ECM in human breast lesions. Specimens collected 18 women undergoing lumpectomy or mastectomy were evaluated with LSM. Because collagen is the key protein associated with ECM stiffness, G*(ω) maps obtained from LSM were compared with collagen content measured by second harmonic generation (SHG) microscopy. Regions of low G*(ω), identified by LSM, corresponded to low-intensity SHG signal and adipose tissue. Likewise, regions with high G*(ω) in LSM images matched high intensity SHG signal caused by desmoplastic collagen accumulation. Quantitative regression analysis demonstrated a strong, statistically significant correlation between G*(ω) and SHG signal intensity (R=0.66 p< 0.01). These findings highlight the capability of LSM for quantifying the ECM micro-mechanics, potentially providing important insights into the biomechanical regulators of breast cancer progression.

Dynamic optical coherence elastography (OCE) techniques have rapidly emerged as a noninvasive way to characterize the biomechanical properties of tissue. However, clinical applications of the majority of these techniques have been unfeasible due to the extended acquisition time because of multiple temporal OCT acquisitions (M-B mode). Moreover, multiple excitations, large datasets, and prolonged laser exposure prohibit their translation to the clinic, where patient discomfort and safety are critical criteria. Here, we demonstrate the feasibility of noncontact true kilohertz frame-rate dynamic optical coherence elastography by directly imaging a focused air-pulse induced elastic wave with a home-built phase-sensitive OCE system. The OCE system was based on a 4X buffered Fourier Domain Mode Locked swept source laser with an A-scan rate of ~1.5 MHz, and imaged the elastic wave propagation at a frame rate of ~7.3 kHz. Because the elastic wave directly imaged, only a single excitation was utilized for one line scan measurement. Rather than acquiring multiple temporal scans at successive spatial locations as with previous techniques, here, successive B-scans were acquired over the measurement region (B-M mode). Preliminary measurements were taken on tissue-mimicking agar phantoms of various concentrations, and the results showed good agreement with uniaxial mechanical compression testing. Then, the elasticity of an in situ porcine cornea in the whole eye-globe configuration at various intraocular pressures was measured. The results showed that this technique can acquire a depth-resolved elastogram in milliseconds. Furthermore, the ultra-fast acquisition ensured that the laser safety exposure limit for the cornea was not exceeded.

Optical Coherence Elastography (OCE) is a non-invasive testing modality that maps the mechanical property of soft tissues with high sensitivity and spatial resolution using phase-sensitive optical coherence tomography (PhS-OCT). Shear wave OCE (SW-OCE) is a leading technique that relies on the speed of propagating shear waves to provide a quantitative elastography. Previous shear wave imaging OCT techniques are based on repeated M-B scans, which have several drawbacks such as long acquisition time and repeated wave stimulations. Recent developments of Fourier domain mode-locked high-speed swept-source OCT system has enabled enough speed to perform KHz B-scan rate OCT imaging. Here we propose ultra-high speed, single shot shear wave imaging to capture single-shot transient shear wave propagation to perform SW-OCE. The frame rate of shear wave imaging is 16 kHz, at A-line rate of ~1.62 MHz, which allows the detection of high-frequency shear wave of up to 8 kHz. The shear wave is generated photothermal-acoustically, by ultra-violet pulsed laser, which requires no contact to OCE subjects, while launching high frequency shear waves that carries rich localized elasticity information. The image acquisition and processing can be performed at video-rate, which enables real-time 3D elastography. SW-OCE measurements are demonstrated on tissue-mimicking phantoms and porcine ocular tissue. This approach opens up the feasibility to perform real-time 3D SW-OCE in clinical applications, to obtain high-resolution localized quantitative measurement of tissue biomechanical property.

An optical coherence tomography (OCT) system with an A-scan rate of 20 kHz was developed for measuring the biomechanical properties of human finger-pad skin. Such an OCT system operates at a center wavelength of 890 nm with a spectral bandwidth of 150 nm resulting in a very good axial resolution of 2.6 μm. The measured sensitivity and sensitivity roll-off of the system were ~93 dB and ~6 dB mm^-1, respectively. Elastographic B-scan images of the human finger-pad skin were constructed by using 1000 A-scans. Deformations of the human finger-pad before and after sliding, while pressed against a transparent optical glass plate under the action of 0.5-2 N force, were examined both at the surface and sub-surface. Biomechanical properties, i.e., deformation of the skin, finger-pad/glass interface contact area were studied.

Optical Coherence Elastography (OCE) is a rapidly developing technique for assessing tissue biomechanical properties. This study demonstrates the first use of the Lorentz force to induce elastic waves within tissue to quantify the elasticity of tissue in combination with a phase-sensitive OCE system at ~1.5 million A-scans per second. The feasibility of this technique was tested on tissue-mimicking agar phantoms of various concentrations. The results as assessed by OCE were in good agreement with standard mechanical testing of the samples. After the preliminary experiments, the stiffness of porcine liver was examined. The results demonstrate that Lorentz force MHz OCE can be applied to study the elasticity of biological tissue effectively and has the potential for clinical applications due to rapid excitation and imaging.

Quantitative probing of the mechanical properties of scattering media by Brillouin spectroscopy is an emerging field of research. At present, Brillouin spectrometers typically detect spontaneous Brillouin backscattered signals from the sample using setups that comprise virtually imaged phased arrays (VIPAs) cascaded in cross-axis configuration or heated molecular absorption cells prior to the VIPA. These experimental arrangements are necessary in order to significantly suppress the strong elastic scattering background from the medium. In this talk, we present a different approach for Brillouin spectroscopy of scattering matter based on stimulated Brillouin scattering (SBS) amplification. Unlike spontaneous Brillouin scattering, SBS amplification does not show elastic scattering background due to the resonant nature of the amplification process, thereby providing excellent spectral contrast. We demonstrate that the use of two continuous-wave distributed feedback lasers at 780 nm in a counter-propagating SBS amplifier geometry is useful for acquiring high signal-to-noise ratio SBS spectra of Intralipid solutions at concentrations that yield up to ~3 scattering events for photons propagating through the sample. Potential applications of SBS spectroscopy in mechanical characterization of thin tissue sections and biopolymers will be discussed.

Brillouin spectroscopy allows non-invasive measurement of the mechanical properties of a sample by measuring the spectra of acoustically induced light scattering therein, and thus has been widely investigated for biomedical application. Recently, the development of fast Brillouin spectrometry based on virtually-imaged phased array (VIPA) has made in-situ measurement of biomedical sample possible. However, one limitation of current Brillouin technique is the low spectral extinction, which limits the measurement to nearly transparent sample. In order to measure turbid sample, multistage VIPA can be cascaded to gain spectral extinction. For example, spectral extinction of ~80 dB was achieved using three-stage VIPA; however, this approach significantly sacrificed measurement throughput. In this work, we develop a novel spectrometer that achieves high extinction without significant signal loss. To achieve this goal, we combine a two-stage VIPA spectrometer with a triple-pass Fabry-Perot interferometer. The triple-pass Fabry-Perot interferometer acts as a band-pass filter with ~3 GHz bandwidth and ~35-dB spectral extinction. Therefore, the overall extinction of this spectrometer greatly surpasses 80 dB with only ~20% excess loss. We demonstrated the performance of this spectrometer measuring background-free Brillouin spectra from Intralipid solutions and within chicken tissue.

Viscoelastic properties of living cells are often directly related to the cell types and their physiological conditions. Unfortunately, all the currently existing methods for analysis of viscoelastic properties of cells, such as micropipette aspiration, atomic force microscopy and optical tweezers are intrinsically slow, limiting their applicability to study large population of cells, which are often needed for either fundamental or clinical studies. In this report, by incorporating the concept of impulsive stimulated Brillouin scattering (ISBS), we report a Brillouin Imaging and Sensing system via Time-Resolved Optical (BISTRO) measurements. We will prove the principle of the BISTRO system by presenting example microscopic measurements and flow/cell cytometry results [1].

In Brillouin scattering imaging, rejection of background noise due to elastic scattering and reflections from optical components is crucial. This is because Brillouin signal is weak, and the signal frequency shift compared with source laser line is very small. Therefore the line of interest is very easy to be contaminated. Whereas physical blocking of undesired component in a dispersed spectrum is used, to filter out background optically provides better stability. Conventional optical filter techniques, such as dielectric-stack filters, holographic volume filters, Lyot fitlers etc. normally have a stopband-width (bandstop filter) or edge-width (edge filter) ranging from a few nanometers to tens of nanometers despite high rejection rate. They cannot be implemented in Brillouin imaging due to the small wavelength shift (< 1 pm). We report a Fabry-Perot etalon-based notch filter for background cleaning in Brillouin imaging. The notch filer takes advantage of multiple reflections of the light beam with a Fabry-Perot etalon to achieve high rejection with narrow bandwidth. The theoretical rejection rate is multiple time of the rejection of one reflection of the etalon. We demonstrated a laser line suppression of > 40 dB while with < 40% power loss with experiments. Width of the stopband at -30 dB rejection level is ~ 1 GHz. This method is not wavelength specific. One etalon may be implanted to a wide spectrum of laser wavelengths. Furthermore, it does not require heating as gaseous notch filters. Our method can also be implemented to Raman scattering, fluorescent imaging and other imaging techniques in which line of interest is close to the laser source.

Spontaneous Brillouin scattering is an inelastic scattering process arising from inherent thermal density fluctuations, or acoustic phonons, propagating in a medium. Over the last few years, Brillouin spectroscopy has shown great potential to become a reliable non-invasive diagnostic tool due to its unique capability of retrieving viscoelastic properties of materials such as strain and stiffness. The detection of the weak scattered light, in addition to the resolution of the Brillouin peaks (typically shifted by few GHz from the central peak) represent one of the greatest challenges in Brillouin. The recent development of high sensitivity CCD cameras has brought Brillouin spectroscopy from a point sampling technique to a new imaging modality. Furthermore, the application of Virtually Imaged Phased Array (VIPA) etalons has dramatically reduced insertion loss simultaneously allowing fast (<1s) collection of the entire spectrum. Hitherto Brillouin microscopy has been shown the ability to provide unique stiffness maps of biological samples, such as the human lens, in a non-destructive manner. In this work, we present results obtained using our Brillouin microscope to map the stiffness variations in the walls of blood vessels in particular when atherosclerotic plaques are formed. The stiffness of the membrane that covers the plaques is critical in developing acute myocardial infarction yet it is not currently possible to credibly assess its stiffness due to lack of suitable methods.

This Conference Presentation was recorded at SPIE Phonics West 2016 held in San Francisco, California, United States.

Changes in tissue biomechanical properties often signify the onset and progression of diseases, such as in determining the vulnerability of atherosclerotic plaques. Acoustic radiation force optical coherence elastography (ARF-OCE) has been used in the detection of tissue elasticity to obtain high-resolution elasticity maps. We have developed a probe-based ARF-OCE technology that utilizes a miniature 10 MHz ring ultrasonic transducer for excitation and Doppler optical coherence tomography (OCT) for detection. The transducer has a small hole in the center for the OCT light to propagate through. This allows for a confocal stress field and light detection within a small region for high sensitivity and localized excitation. This device is a front-facing probe that is only 3.5 mm in diameter and it is the smallest ARF-OCE catheter to the best of our knowledge. We have tested the feasibility of the probe by measuring the point displacement of an agarose tissue-mimicking phantom using different ARF excitation voltages. Small displacement values ranging from 30 nm to 90 nm have been detected and are shown to be directly proportional to the excitation voltage as expected. We are currently working on obtaining 2D images using a scanning mechanism. We will be testing to capture 2D elastograms of phantoms to further verify feasibility, and eventually characterize the mechanical properties of cardiovascular tissue. With its high portability and sensitivity, this novel technology can be applied to the diagnosis and characterization of vulnerable atherosclerotic plaques.

Acute Glomerulonephritis caused by anti-glomerular basement membrane disease has a high mortality due to delayed diagnosis. Thus, an accurate and early diagnosis is critical for preserving renal function. Currently, blood, urine, and tissue-based diagnoses can be time consuming, while ultrasound and CT imaging have relatively low spatial resolution. Optical coherence tomography (OCT) is a noninvasive imaging technique that provides superior spatial resolution (micron scale) as compared to ultrasound and CT. Pathological changes in tissue properties can be detected based on the optical metrics analyzed from the OCT signal, such as optical attenuation and speckle variance. Moreover, OCT does not rely on ionizing radiation as with CT imaging. In addition to structural changes, the elasticity of the kidney can significantly change due to nephritis. In this work, we utilized OCT to detect the difference in tissue properties between healthy and nephritic murine kidneys. Although OCT imaging could identify the diseased tissue, classification accuracy using only optical metrics was clinically inadequate. By combining optical metrics with elasticity, the classification accuracy improved from 76% to 95%. These results show that OCT combined with OCE can be potentially useful for nephritis detection.

Optical coherence elastography (OCE) asesses the mechanical properties of samples by applying a mechanical stimulation and detecting the resulting sample displacement using optical coherence tomography (OCT). OCE methods which utilise the phase of the OCT signal offer the potential to detect displacements on the sub-nanometre scale. However, the displacement sensitivity achieveable is directly related to the signal-to-noise ratio and phase stability of the underlying OCT system. Furthermore, the estimation of Doppler angle is imperative in accurately measuring the sample displacement. This work evaluates the contributions of each of these parameters for quantitative assessment of mechanical properties using phase-sensitive spectral domain OCT.

We report a novel hybrid method of robust strain mapping in compressional optical coherence elastography using combined phase measurements on sub-wavelength-scale and cumulative pixel-scale displacement tracking. This hybrid nature significantly extends the range of measurable displacements and strains in comparison with conventional direct phase-resolved measurements. As a result, the proposed strain-mapping method exhibits significantly increased robustness with respect to both additive noise and decorrelation noise produced by displacements and strains. The main advantages of the proposed approach are illustrated by numerical simulations. Experimental examples of obtained strain maps for phantoms and real biological tissues are also presented.

The diaphragm, composed of skeletal muscle, plays an important role in respiration through its dynamic contraction. Genetic and molecular studies of the biomechanics of mouse diaphragm can provide great insights into an improved understanding and potential treatment of the disorders that lead to diaphragm dysfunction (i.e. muscular dystrophy). However, due to the small tissue size, mechanical assessment of mouse diaphragm tissue under its proper physiological conditions has been challenging. Here, we present the application of noncontact optical coherence elastography (OCE) for quantitative elastic characterization of ex vivo mouse diaphragm. Phase-sensitive optical coherence tomography was combined with a focused air-puff system to capture and measure the elastic wave propagation from tissue surface. Experiments were performed on wildtype and dystrophic mouse diaphragm tissues containing different levels of fibrosis. The OCE measurements of elastic wave propagation were conducted along both the longitudinal and transverse axis of the muscle fibers. Cross-correlation of the temporal displacement profiles from different spatial locations was utilized to obtain the propagation time delay, which was used to calculate the wave group velocity and to further quantify the tissue Young’s modulus. Prior to and after OCE assessment, peak tetanic force was measured to monitor viability of the tissue during the elasticity measurements. Our experimental results indicate a positive correlation between fibrosis level and tissue stiffness, suggesting this elastic-wave-based OCE method could be a useful tool to monitor mechanical properties of skeletal muscle under physiological and pathological conditions.

Biomechanical properties of tissues have been utilized for disease detection, diagnosis, and progression, however they have not been extensively utilized for therapy dosimetry. Magnetic hyperthermia aims to kill cells and ablate tumors using magnetic nanoparticles (MNPs) either injected in or targeted to tumors. Upon application of an appropriate AC magnetic field, MNPs can heat target tissue while sparing non-targeted healthy tissue. However, a sensitive monitoring technique for the dose of magnetic hyperthermia is needed to prevent over-treatment and collateral injury. During hyperthermia treatments, the viscoelastic properties of tissues are altered due to protein denaturation, coagulation, and tissue dehydration, making these properties candidates for dosimetry. Magnetomotive optical coherence elastography (MM-OCE) utilizes MNPs as internal force transducers to probe the biomechanical properties of tissues. Therefore, we aim to evaluate the hyperthermia dose based on the elastic changes revealed by MM-OCE. In this study, MNPs embedded in tissues were utilized for both hyperthermia and MM-OCE measurements. Tissue temperature and elastic modulus were obtained, where the elastic modulus was extracted from the resonance frequency detected by MM-OCE. Results showed a correlation between stiffness and temperature change following treatment. To investigate the thermal-dose-dependent changes, intervals of hyperthermia treatment were repeatedly performed on the same tissue sequentially, interspersed with MM-OCE. With increasing times of treatment, tissue stiffness increased, while temperature rise remained relatively constant. These results suggest that MM-OCE may potentially identify reversible and irreversible tissue changes during thermal therapy, supporting the use of MM-OCE for dosimetric control of hyperthermia in future applications.

A number of disease conditions including coronary atherosclerosis, peripheral artery disease and gastro-intestinal malignancies are associated with alterations in tissue mechanical properties. Laser speckle rheology (LSR) has been demonstrated to provide important information on tissue mechanical properties by analyzing the time scale of temporal speckle intensity fluctuations, which serves as an index of tissue viscoelasticity. In order to measure the mechanical properties of luminal organs in vivo, LSR must be conducted via a miniature endoscope or catheter. Here we demonstrate the capability of an omni-directional LSR catheter to quantify tissue mechanical properties over the entire luminal circumference without the need for rotational motion. Retracting the catheter using a motor-drive assembly enables the reconstruction of cylindrical maps of tissue mechanical properties. The performance of the LSR catheter is tested using a luminal phantom with mechanical moduli that vary in both circumferential and longitudinal directions. 2D cylindrical maps of phantom viscoelastic properties are reconstructed over four quadrants of the coronary circumference simultaneously during catheter pullback. The reconstructed cylindrical maps of the decorrelation time constants easily distinguish the different gel components of the phantom with different viscoelastic moduli. The average values of decorrelation times calculated for each gel component of the phantom show a strong correspondence with the viscoelastic moduli measured via standard mechanical rheometry. These results highlight the capability for cylindrical mapping of tissue viscoelastic properties using LSR in luminal organs using a miniature catheter, thus opening the opportunity for improved diagnosis of several disease conditions.

In this study we have evaluated the elastic anisotropy of porcine corneas with increasing intraocular pressures (IOPs) using a noncontact optical coherence elastography (OCE) technique. A focused air-pulse induced low amplitude (≤10 μm) elastic waves in fresh porcine corneas (n=9) in situ in the whole eye-globe configuration. A phase-stabilized swept source optical coherence elastography (PhS-SSOCE) system imaged the propagation of the elastic wave in different stepped radial directions. A closed-loop feedback system was utilized to artificially manipulate the IOP, and OCE measurements were repeated while the IOP was increased in 5 mmHg increments from 15 to 30 mmHg. The OCE measurements demonstrated that the elastic anisotropy of the cornea became more pronounced at higher IOPs, and that there were distinct radial angles of higher and lower stiffness. The presented noncontact OCE method was capable of detecting and assessing the corneal elastic anisotropy as a function of IOP. Due to the noninvasive nature and small amplitude of the elastic wave, this method may be able to provide additional information about corneal health and integrity in vivo.

Here, we present the first catheter-based optical coherence elasticity measurement using a newly developed super fast intravascular optical coherence tomography (OCT) system. The system is based on a 1.5 MHz Fourier Domain Mode Locked laser and a 1.2 mm outer diameter motorized catheter. To detect the local elastic properties, the micro-motor is programmed to actuate the laser beam in a “step-by-step” mode at 1 revolution per second; which can potentially be increased to > 10 revolutions/s. The beam is scanned in a limited number (up to 50) of angular steps, at each of which the beam position is held stable. When the laser beam is stable, the phase difference across a variable number of A-lines can be computed to assess the elastic displacement. Choosing a proper window delay, local elastic tissue displacement and strain can be quantified based on the phase shift. We conducted ex-vivo experiments with a cylindrical phantom where the elastic property changes at different angular positions. A syringe pump was used to generate variable pressure loading, which is synchronized to the motor driving signal. The experimental results show that the elastic displacements are detected to be different at different angular positions. The results of elastic properties detection in human artery will also be demonstrated.

What stem cells become depends in part on what they 'mechanically' feel around them. Soft tissues such as fat bear little physical stress and have less abundant structural protein, whereas stiffer tissues like muscle and bone sustain high stress and have a relative abundance of structural proteins. We have begun to uncover systematic relationships between such tissue properties and differentiation processes, having first shown that a soft matrix helps specify soft tissue lineages of stem cells while a stiff matrix helps specify stiff tissue lineages of stem cells. These general principles seem to apply to normal stem cells and perhaps apply or go awry for cancer stem cells.

It is now well recognized that a host of imaging modalities (a list that includes Ultrasound, MRI, Optical Coherence Tomography, and optical microscopy) can be used to “watch” tissue as it deforms in response to an internal or external excitation. The result is a detailed map of the deformation field in the interior of the tissue. This deformation field can be used in conjunction with a material mechanical response to determine the spatial distribution of material properties of the tissue by solving an inverse problem. Images of material properties thus obtained can be used to quantify the health of the tissue. Recently, they have been used to detect, diagnose and monitor cancerous lesions, detect vulnerable plaque in arteries, diagnose liver cirrhosis, and possibly detect the onset of Alzheimer’s disease. In this talk I will describe the mathematical and computational aspects of solving this class of inverse problems, and their applications in biology and medicine. In particular, I will discuss the well-posedness of these problems and quantify the amount of displacement data necessary to obtain a unique property distribution. I will describe an efficient algorithm for solving the resulting inverse problem. I will also describe some recent developments based on Bayesian inference in estimating the variance in the estimates of material properties. I will conclude with the applications of these techniques in diagnosing breast cancer and in characterizing the mechanical properties of cells with sub-cellular resolution.

Wave models, which have been utilized in the past to reconstruct corneal biomechanical properties based on the propagation of an elastic wave, were often developed assuming a thin-plate geometry. However, the curvature and thickness of the cornea are not considered when utilizing these models. In this work, optical coherence elastography (OCE) experiments were conducted on tissue-mimicking agar phantoms and contact lenses along with finite element (FE) modeling of four kinds of cornea-like structures to understand the effects of curvature and thickness on the group velocity of an elastic wave. As the radius of curvature increased from 19.1 to 47.7 mm, the group velocity of the elastic wave obtained by both FE and OCE from a spherical shell section model decreased from ~2.8 m/s to ~2.2 m/s. When the thickness of the agar phantom increased from 1.9 mm to 5.6 mm, the elastic wave velocity increased from ~3.0 m/s to ~4.1 m/s. Both the FE and OCE results show that the group velocity of the elastic wave decreased with radius of curvature but increased with thickness. Therefore, the curvature and thickness must be considered when developing accurate wave models for quantifying biomechanical properties of the cornea.

Optical Coherence Elastography (OCE) is a widely investigated noninvasive technique for estimating the mechanical properties of tissue. In particular, vibrational OCE methods aim to estimate the shear wave velocity generated by an external stimulus in order to calculate the elastic modulus of tissue. In this study, we compare the performance of five acquisition and processing techniques for estimating the shear wave speed in simulations and experiments using tissue-mimicking phantoms. Accuracy, contrast-to-noise ratio, and resolution are measured for all cases. The first two techniques make the use of one piezoelectric actuator for generating a continuous shear wave propagation (SWP) and a tone-burst propagation (TBP) of 400 Hz over the gelatin phantom. The other techniques make use of one additional actuator located on the opposite side of the region of interest in order to create an interference pattern. When both actuators have the same frequency, a standing wave (SW) pattern is generated. Otherwise, when there is a frequency difference df between both actuators, a crawling wave (CrW) pattern is generated and propagates with less speed than a shear wave, which makes it suitable for being detected by the 2D cross-sectional OCE imaging. If df is not small compared to the operational frequency, the CrW travels faster and a sampled version of it (SCrW) is acquired by the system. Preliminary results suggest that TBP (error < 4.1%) and SWP (error < 6%) techniques are more accurate when compared to mechanical measurement test results.

Various types of waves are produced when a harmonic force is applied to a semi-infinite half space elastic medium. In particular, surface waves are perturbations with transverse and longitudinal components of displacement that propagate in the boundary region at the surface of the elastic solid. Shear wave speed estimation is the standard for characterizing elastic properties of tissue in elastography; however, the penetration depth of Optical Coherence Tomography (OCT) is typically measured in millimeters constraining the measurement region of interest to be near the surface. Plane harmonic Rayleigh waves propagate in solid-vacuum interfaces while Scholte waves exist in solid-fluid interfaces. Theoretically, for an elastic solid with a Poisson’s ratio close to 0.5, the ratio of the Rayleigh to shear wave speed is 95%, and 84% for the Scholte to shear wave. Our study demonstrates the evidence of Rayleigh waves propagating in the solid-air boundary of tissue-mimicking elastic phantoms. Sinusoidal tone-bursts of 400Hz and 1000 Hz were excited over the phantom by using a piezoelectric actuator. The wave propagation was detected with a phase-sensitive OCT system, and its speed was measured by tracking the most prominent peak of the tone in time and space. Similarly, this same experiment was repeated with a water interface. In order to obtain the shear wave speed in the material, mechanical compression tests were conducted in samples of the same phantom. A 93.9% Rayleigh-shear and 82.4% Scholte-Shear speed ratio were measured during experiments which are in agreement with theoretical results.

We have derived an axially symmetric three-dimensional analytical solution for thermoelastic deformations and stresses in a layered medium irradiated by a laser beam. The solution was obtained for Gaussian radial temperature profile on the upper surface of the elastic layer, in the assumption that temperature decreases exponentially with depth. The developed theoretical model was used to calculate distributions of laser-induced deformations and displacements in a medium containing single layer over a half-space. The influence of the shear elastic properties of the layer and half-space on the stress and strain distributions was evaluated. It was shown that tissue response depends significantly on the elastic contrast between the layer and the half-space. The proposed solution could be used in photomechanical models of laser ablation of inhomogeneous materials and tissues.

Quantitative elasticity imaging, which retrieves elastic modulus maps from tissue, is preferred to qualitative strain imaging for acquiring system- and operator-independent images and longitudinal and multi-site diagnoses. Quantitative elasticity imaging has already been demonstrated in optical elastography by relating surface-acoustic and shear wave speed to Young’s modulus via a simple algebraic relationship. Such approaches assume largely homogeneous samples and neglect the effect of boundary conditions. We present a general approach to quantitative elasticity imaging based upon the solution of the inverse elasticity problem using an iterative technique and apply it to compression optical coherence elastography. The inverse problem is one of finding the distribution of Young’s modulus within a sample, that in response to an applied load, and a given displacement and traction boundary conditions, can produce a displacement field matching one measured in experiment. Key to our solution of the inverse elasticity problem is the use of the adjoint equations that allow the very efficient evaluation of the gradient of the objective function to be minimized with respect to the unknown values of Young’s modulus within the sample. Although we present the approach for the case of linear elastic, isotropic, incompressible solids, this method can be employed for arbitrarily complex mechanical models. We present the details of the method and quantitative elastograms of phantoms and tissues. We demonstrate that by using the inverse approach, we can decouple the artefacts produced by mechanical tissue heterogeneity from the true distribution of Young’s modulus, which are often evident in techniques that employ first-order algebraic relationships.

Accurate quantification of forces, applied to, or generated by, tissue, is key to understanding many biomechanical processes, fabricating engineered tissues, and diagnosing diseases. Many techniques have been employed to measure forces; in particular, tactile imaging – developed to spatially map palpation-mimicking forces – has shown potential in improving the diagnosis of cancer on the macro-scale. However, tactile imaging often involves the use of discrete force sensors, such as capacitive or piezoelectric sensors, whose spatial resolution is often limited to 1-2 mm. Our group has previously presented a type of tactile imaging, termed optical palpation, in which the change in thickness of a compliant layer in contact with tissue is measured using optical coherence tomography, and surface forces are extracted, with a micro-scale spatial resolution, using a one-dimensional spring model. We have also recently combined optical palpation with compression optical coherence elastography (OCE) to quantify stiffness. A main limitation of this work, however, is that a one-dimensional spring model is insufficient in describing the deformation of mechanically heterogeneous tissue with uneven boundaries, generating significant inaccuracies in measured forces. Here, we present a computational, finite-element method, which we term computational optical palpation. In this technique, by knowing the non-linear mechanical properties of the layer, and from only the axial component of displacement measured by phase-sensitive OCE, we can estimate, not only the axial forces, but the three-dimensional traction forces at the layer-tissue interface. We use a non-linear, three-dimensional model of deformation, which greatly increases the ability to accurately measure force and stiffness in complex tissues.

Object of study: A study of the biomechanical characteristics of the human heart ventricles was performed. 80 hearts were extracted during autopsy of 80 corpses of adults (40 women and 40 men) aged 31-70 years. The samples were investigated in compliance with the recommendations of the ethics committee.

Methods: Tension and compression tests were performed with help of the uniaxial testing machine Instron 5944. Cardiometry was also performed.

Results: In this work, techniques for human heart ventricle wall biomechanical properties estimation were developed. Regularities of age and gender variability in deformative and strength properties of the right and left ventricle walls were found. These properties were characterized by a smooth growth of myocardial tissue stiffness and resistivity at a relatively low strain against reduction in their strength and elasticity from 31-40 to 61-70 years. It was found that tissue of the left ventricle at 61-70 years had a lower stretchability and strength compared with tissues of the right ventricle and septum. These data expands understanding of the morphological organization of the heart ventricles, which is very important for the development of personalized medicine. Taking into account individual, age and gender differences of the heart ventricle tissue biomechanical characteristics allows to rationally choosing the type of patching materials during reconstructive operations on heart.

Object of study: The research is aimed at development of personalized medical treatment. Algorithm was developed for patient-specific surgical interventions of the cardiovascular system pathologies.

Methods: Geometrical models of the biological objects and initial and boundary conditions were realized by medical diagnostic data of the specific patient. Mechanical and histomorphological parameters were obtained with the help mechanical experiments on universal testing machine. Computer modeling of the studied processes was conducted with the help of the finite element method.

Results: Results of the numerical simulation allowed evaluating the physiological processes in the studied object in normal state, in presence of different pathologies and after different types of surgical procedures.

It is important to measure embryonic heart myocardial wall strain and strain rate for understanding the mechanisms of embryonic heart development. Optical coherence tomography (OCT) can provide depth resolved images with high spatial and temporal resolution, which makes it have the potential to reveal the complex myocardial activity in the early stage embryonic heart. We develop a novel method to measure strain in embryonic chick heart based on spectral domain OCT images and subsequent image processing. We perform 4D(x,y,z,t) scanning on the outflow tract (OFT) of chick embryonic hearts in HH18 stage (~3 days of incubation). Only one image sequence acquired at the special position is selected based on the Doppler blood flow information where the probe beam penetrates through the OFT perpendicularly. For each image of the selected sequence, the cross-section of the myocardial wall can be approximated as an annulus. The OFT is segmented with a semi-automatic boundary detection algorithm, thus the area and mean circumference of the annular myocardial wall can be achieved. The myocardial wall thickness was calculated using the area divided by the mean circumference, and then the strain was obtained. The results demonstrate that OCT can be a useful tool to describe the biomechanical characteristics of the embryonic heart.

Bio-mechanical properties of the human skin deformed by external forces at difference skin/material interfaces attract much attention in medical research. For instance, such properties are important design factors when one designs a healthcare device, i.e., the device might be applied directly at skin/device interfaces. In this paper, we investigated the bio-mechanical properties, i.e., surface strain, morphological changes of the skin layers, etc., of the human finger-pad and forearm skin as a function of applied pressure by utilizing two non-invasive techniques, i.e., optical coherence tomography (OCT) and digital image correlation (DIC). Skin deformation results of the human finger-pad and forearm skin were obtained while pressed against a transparent optical glass plate under the action of 0.5-24 N force and stretching naturally from 90° flexion to 180° full extension respectively. The obtained OCT images showed the deformation results beneath the skin surface, however, DIC images gave overall information of strain at the surface.

Our group previously introduced Polarized Spatial Frequency Domain Imaging (PSFDI), a wide-field, reflectance imaging technique which we used to empirically map fiber direction in porcine pulmonary heart valve leaflets (PHVL) without optical clearing or physical sectioning of the sample. Presented is an extended analysis of our PSFDI results using an inverse Mueller matrix model of polarized light scattering that allows additional maps of fiber orientation distribution, along with instrumentation permitting increased imaging speed for dynamic PHVL fiber measurements.

We imaged electrospun fiber phantoms with PSFDI, and then compared these measurements to SEM data collected for the same phantoms. PHVL was then imaged and compared to results of the same leaflets optically cleared and imaged with small angle light scattering (SALS). The static PHVL images showed distinct regional variance of fiber orientation distribution, matching our SALS results. We used our improved imaging speed to observe bovine tendon subjected to dynamic loading using a biaxial stretching device. Our dynamic imaging experiment showed trackable changes in the fiber microstructure of biological tissue under loading. Our new PSFDI analysis model and instrumentation allows characterization of fiber structure within heart valve tissues (as validated with SALS measurements), along with imaging of dynamic fiber remodeling. The experimental data will be used as inputs to our constitutive models of PHVL tissue to fully characterize these tissues' elastic behavior, and has immediate application in determining the mechanisms of structural and functional failure in PHVLs used as bio-prosthetic implants.

Quantitative elastography is a power technique to detect and analyze the changes in biomedical properties of tissues in normal and pathological states. In this study, two noncontact elastography techniques, laser Michelson vibrometry (LMV) and optical coherence elastography (OCE), were utilized to quantify the Young’s modulus of tissue-mimicking agar phantoms of various concentrations. Low-amplitude (micrometer scale) elastic waves were induced by a focused air-pulse delivery system and imaged by the respective systems. The Young’s modulus as assessed by both elastographic techniques was similar and was compared to the stiffness as measured by uniaxial mechanical testing. The results show that both techniques accurately quantified the elasticity. OCE can provide absolute elastic wave temporal profile, depth-resolved measurement and superior displacement sensitivity compared to LMV, but LMV is significantly cheaper (10X) and easier to implement than OCE.

Incomplete excision of tumour margins is a major issue in breast-conserving surgery. Currently 20 – 60% of cases require a second surgical procedure required as a result of cancer recurrence. A number of techniques have been proposed to assess margin status, including frozen section analysis and imprint cytology. However, the recurrence rate after using these techniques remains very high. Over the last several years, our group has been developing optical coherence elastography (OCE) as a tool for the intraoperative assessment of tumour margins in breast cancer. We have reported a feasibility study on 65 ex vivo samples from patients undergoing mastectomy or wide local excision demonstrates the potential of OCE in differentiating benign from malignant tissue. In this study, malignant tissue was readily distinguished from surrounding relative tissue by a distinctive heterogeneous pattern in micro-elastograms. To date the largest field of view for a micro-elastogram is 20 x 20mm, however, lumpectomy samples are typically ~50 x 50 x 30mm. For OCE to progress as a useful clinical tool, elastograms must be acquired over larger areas to allow a greater portion of the surface area of lumpectomies to be assessed. Here, we propose a wide-field OCE scanner that utilizes a piezoelectric transducer with an internal diameter of 65mm. In this approach partially overlapped elastograms are stitched together forming a mosaic with overall dimensions of 50 x 50mm in a total acquisition time of 15 - 30 minutes. We present results using this approach on both tissue-mimicking phantoms and tissue, and discuss prospects for shorter acquisitions times.

Photothermal OCT has been emerged to contrast absorbers in biological tissues. The tissues response to photothermal excitation as change of thermal strain and refractive index. To resolve the depth of absorption agents, the measurements of the local thermal strain change and local refractive index change due to photothermal effect is required. In this study, we developed photothermal OCT for depth-resolved absorption contrast imaging. The phase-resolved OCT can measure the axial strain change and local refractive index change as local optical path length change. A swept-source OCT system is used with a wavelength swept laser at 1310 nm with a scanning rate of 50 kHz. The sensitivity of 110 dB is achieved. At the sample arm, the excitation beam from a fiber-coupled laser diode of 406 nm wavelength is combined with the OCT probe beam co-linearly. The slowly modulated excitation beam around 300 Hz illuminate biological tissues. M-mode scan is applied during one-period modulation duration. The local optical path length change is measured by temporal and axial phase difference. The theoretical prediction of the photothermal response is derived and in good agreement with experimental results. In the case of slow modulation, the delay of photothermal response can be neglected. The local path length changes are averaged over the half period of the excitation modulation, and then demodulated. This method exhibits 3-dB gain in the sensitivity of the local optical path length change measurement over the direct Fourier transform method. In vivo human skin imaging of endogenous absorption agent will be demonstrated.

Phase-sensitive optical coherence tomography (PhS-OCT) can be utilized for quantitative shear-wave elastography using speckle tracking. However, current approaches cannot directly reconstruct elastic properties in speckle-less or speckle-free regions, for example within the crystalline lens in ophthalmology. Investigating the elasticity of the crystalline lens could improve understanding and help manage presbyopia-related pathologies that change biomechanical properties. We propose to reconstruct the elastic properties in speckle-less regions by sequentially launching shear waves with moving acoustic radiation force (mARF), and then detecting the displacement at a specific speckle-generating position, or limited set of positions, with PhS-OCT. A linear ultrasound array (with a center frequency of 5 MHz) interfaced with a programmable imaging system was designed to launch shear waves by mARF. Acoustic sources were electronically translated to launch shear waves at laterally shifted positions, where displacements were detected by speckle tracking images produced by PhS-OCT operating in M-B mode with a 125-kHz A-line rate. Local displacements were calculated and stitched together sequentially based on the distance between the acoustic source and the detection beam. Shear wave speed, and the associated elasticity map, were then reconstructed based on a time-of-flight algorithm. In this study, moving-source shear wave elasticity imaging (SWEI) can highlight a stiff inclusion within an otherwise homogeneous phantom but with a CNR increased by 3.15 dB compared to a similar image reconstructed with moving-detector SWEI. Partial speckle-free phantoms were also investigated to demonstrate that the moving-source sequence could reconstruct the elastic properties of speckle-free regions. Results show that harder inclusions within the speckle-free region can be detected, suggesting that this imaging method may be able to detect the elastic properties of the crystalline lens.

Each year, in the Netherlands alone, more than 50.000 percutaneous procedures are performed for treatment or for removal of tissue from possibly diseased organs, of which 30% return non-diagnostic due to erroneous needle targeting, often as a result of non-homogeneity of the penetrated tissue. In this study, we aim to facilitate needle targeting by assessing the tissue in front of the needle based on its mechanical properties. A probe that can identify tissues via real-time measurements of their mechanical properties is placed at the tip of the needle. The probe, actuated by a remote system at the distal part of the needle, employs the bending of a micro-machined cantilever fabricated on top of an optical fiber. The displacement of the cantilever, imposed by pressing a micro-bead (r = 75 µm) glued at the tip of the cantilever against the tissue, is interrogated by Fabry-Pérot interferometry and converted to force acted on the tissue in real-time. The force transducer is able to perform in harsh environments due to its monolithic design and all-optical working principle. Using our setup, load-indentation curves were obtained during needle insertion in several gelatin-based specimens. We demonstrate the ability of our device to detect and quantify layers of varying stiffness and to successfully locate tissue boundaries in animal tissue embedded in gelatin. Furthermore, a diagnostic measurement can be made by quantifying intra-organ tissue stiffness at the needle target location.