Uncertainties of synchrotron microCT-based digital volume correlation bone strain measurements under simulated deformation

Digital Volume Correlation (DVC) is used to measure internal displacements and strains in bone. Recent studies have shown that Synchrotron radiation micro-computed tomography (SR-microCT) can improve the accuracy and precision of DVC. However, only zero-strain or virtually-moved test have been used to quantify the DVC uncertainties, leading to potential underestimation of the measurement errors.In this study, for the first time, the uncertainties of a global DVC approach have been evaluated on repeated SR-microCT scans of bovine cortical bone (voxel size: 1.6 μm), which were virtually deformed for different magnitudes and along different directions.The results showed that systematic and random errors of the normal strain components along the deformation direction were higher than the errors along unstrained directions. The systematic percentage errors were smaller for larger virtual deformations. The random percentage error was in the order of 10% of the virtual deformation. However, higher errors were localized at the boundary of the volumes of interest, perpendicular to the deformation direction. When only the central region of the samples was considered (100 µm layers removed from the borders where the deformation was applied), the errors in the direction of virtual deformation were comparable to the errors in the unstrained directions.In conclusion, the method presented to estimate the uncertainties of DVC is suitable for testing anisotropic specimens as cortical bone. The good agreement between the uncertainties in measurements of strain components obtained with this approach and with the simpler zero-strain-test suggests that the latter is adequate in the tested deformation scenarios.     

High resolution three-dimensional strain mapping of bioprosthetic heart valves using digital image correlation

Transcatheter aortic valve replacement (TAVR) is a safe and effective treatment option for patients deemed at high and intermediate risk for surgical aortic valve replacement. Similar to surgical aortic valves (SAVs), transcatheter aortic valves (TAVs) undergo calcification and mechanical wear over time. However, to date, there have been limited publications on the long-term durability of TAV devices. To assess longevity and mechanical strength of TAVs in comparison to surgical bioprosthetic valves, three-dimensional deformation analysis and strain measurement of the leaflets become an inevitable part of the evaluation. The goal of this study was to measure and compare leaflet displacement and strain of two commonly used TAVs in a side-by-side comparison with a commonly used SAV using a high-resolution digital image correlation (DIC) system. 26-mm Edwards SAPIEN 3, 26-mm Medtronic CoreValve, and 25-mm Carpentier-Edwards PERIMOUNT Magna surgical bioprosthesis were examined in a custom-made valve testing apparatus. A time-varying, spatially uniform pressure was applied to the leaflets at different loading rates. GOM ARAMIS® software was used to map leaflet displacement and strain fields during loading and unloading. High displacement regions were found to be at the leaflet belly region of the three bioprosthetic valves. In addition, the frame of the surgical bioprosthesis was found to be remarkably flexible, in contrary to CoreValve and SAPIEN 3 in which the stent was nearly rigid under a similar loading condition. The experimental DIC measurements can be used to characterize the anisotropic materiel behavior of the bioprosthetic heart valve leaflets and validate heart valve computational simulations.     

In-situ characterization of the uncrimping process of arterial collagen fibers using two-photon confocal microscopy and digital image correlation

Uncrimping of collagen fibers in the arterial wall is an integral process in regulating the macro-level mechanical response of arteries. Uncrimping of collagen fibers leads to a gradual, but significant strain-stiffening response of the artery at physiological pressures and prevents overdistention at elevated pressures. In this study, we imaged adventitial collagen fibers from fresh primate arteries using two-photon excitation microscopy while subjecting the arteries to physiological inflation pressures and axial stretches. The imaging focal plane was fixed at a constant radial location in the adventitial wall by adjusting the focal distance as the arteries inflated, allowing for the continuously monitoring of the uncrimping process of a single region of collagen fibers. Digital image correlation was then applied to the sequential images to assess and correlate the local displacements to manual traces of selected reference fibers and their engagements. We found that the collagen fibers of interest became fully engaged at a luminal pressure of 20 mmHg, this was then followed by rotation of these fibers as the bulk artery continued to dilate. This technique helps to further the understanding of the uncrimping process of collagen fibers under physiological loads, which can aid in the development of more accurate microstructural constitutive models.     

Cell and tissue deformation measurements: Texture correlation with third-order approximation of displacement gradients

Cells remarkably are capable of large deformations during motility and when subjected to mechanical force. Measurement of mechanical deformation (i.e. displacements, strain) is critical to understand functional changes in cells and biological tissues following disease, and to elucidate basic relationships between applied force and cellular biosynthesis. Microscopy-based imaging modalities provide the ability to noninvasively visualize small cell or tissue structures and track their motion over time, often using two-dimensional (2D) digital image (texture) correlation algorithms. For the measurement of complex and nonlinear motion in cells and tissues, implementation of texture correlation algorithms with high order approximations of displacement mapping terms are needed to minimize error. Here, we extend a texture correlation algorithm with up to third-order approximation of displacement mapping terms for the measurement of cell and tissue deformation. We additionally investigate relationships between measurement error and image texture, defined by subset entropy. Displacement measurement error is significantly reduced when the order of displacement mapping terms in the texture correlation algorithm matches or exceeds the order of the deformation observed. Displacement measurement error is also inversely proportional to subset entropy, with well-defined cell and tissue structures leading to high entropy and low error. For cell and tissue studies where complex or nonlinear displacements are expected, texture correlation algorithms with high order terms are required to best characterize the observed deformation.     

Application of the digital volume correlation technique for the measurement of displacement and strain fields in bone: A literature review

Digital volume correlation (DVC) provides experimental measurements of displacements and strains throughout the interior of porous materials such as trabecular bone. It can provide full-field continuum- and tissue-level measurements, desirable for validation of finite element models, by comparing image volumes from subsequent µCT scans of a sample in unloaded and loaded states.     

Abundance estimation of rocky shore invertebrates at small spatial scale by high-resolution digital photography and digital image analysis

We have tested both the usefulness of high-resolution digital photography for data acquisition and digital image analysis, by non-supervised classification and high pass filter, for recognition and abundance estimation of benthic intertidal organisms. These digital tools were compared with visual scan and photo quadrat conventional methods. The comparison was done using 40 quadrats (10×5 cm) randomly selected along a 5-m transect on the rocky shore of the Pemaquid Point, Maine, USA. ANOVA for repeated measures was used to test differences among methods. Monte Carlo simulation analysis was used to explore differences among methods over a large set of data (n=100, 500, 1000 quadrats). Differences among methods were observed when 40 quadrats were used. Tukey multiple comparison test showed that abundance estimation from visual scan, photo quadrat and digital image analysis by high pass filter do not differ significantly among them but differ from non-supervised classification results. Due to its accurate estimation, high pass filter (Prewitt) method was chosen as the most reliable digital method to estimate species abundance. Monte Carlo simulation of visual scan, photo quadrat and high pass filter results showed significant differences when the number of quadrats was larger. These results showed that the combined use of digital photography and digital image analysis techniques for the acquisition and analysis of recorded data is a powerful method for the study of intertidal benthic organisms. Results produced using these techniques were similar than those produced by conventional methods but were obtained in a much-reduced time.     

Applying the digital image correlation method to estimate the mechanical properties of bacterial biofilms subjected to a wall shear stress

A digital image correlation (DIC) method was applied to characterize the mechanical behavior of Pseudomonas aeruginosa biofilms in response to wall shear stress using digital video micrographs taken from biofilm flow cells. The appearance of the biofilm in the transmitted light photomicrographs presented a natural texture which was highly conducive to random encoding for DIC. The displacement fields were calculated for two biofilm specimens. The DIC method concurred with previous analysis showing that biofilms exhibit viscoelastic behavior, but had the advantage over simple length measurements of longitudinal strain that it could precisely measure local strains in length (x) and width (y) within biofilm clusters with a 2 μm resolution as a function of time and wall shear stress. It was concluded that DIC was more accurate at measuring elastic moduli than simple length measurements, but that time-lapse 3D images would enable even more accurate estimates to be performed.     

Temperature-dependent electrical resistivity of the hornet cuticle: Changes induced by colchicine and purine in the diet

Various cuticular areas of the Oriental hornet (Vespa orientalis) were measured for their electrical resistivity within the range of 3–40°C. It was found that: (1) the electrical resistivity is temperature dependent; (2) there are no significant differences in this respect between hornets collected in different regions; (3) there are no differences in the electrical resisitivity between the yellow and brown areas of the cuticle in control hornets; (4) there are however, significant changes in resistance between the yellow and brown strips of hornets fed on purine; (5) hornets fed on colchicine reach a minimum resistance at a higher temperature than do the control hornets; and (6) hornets fed on purine reach a minimum resistance at a lower temperature. It is suggested that the cuticular changes in resistivity at different temperatures reflect the hornets' mechanism of detecting and adjusting to changes in the ambient temperature.     

3D micro-scale deformations of wood in bending: synchrotron radiation muCT data analyzed with digital volume correlation

A micro-scale three-point-bending experiment with a wood specimen was carried out and monitored by synchrotron radiation micro-computed tomography. The full three-dimensional wood structure of the 1.57 × 3.42 × 0.75 mm³ specimen was reconstructed at cellular level in different loading states. Furthermore, the full three-dimensional deformation field of the loaded wood specimen was determined by digital volume correlation, applied to the reconstructed data at successive loading states. Results from two selected regions within the wood specimen are presented as continuous displacement and strain fields in both 2D and 3D. The applied combination of synchrotron radiation micro-computed tomography and digital volume correlation for the deformation analysis of wood under bending stress is a novel application in wood material science. The method offers the potential for the simultaneous observation of structural changes and quantified deformations during in situ micro-mechanical experiments. Moreover, the high spatial resolution allows studying the influence of anatomical features on the fracture behaviour of wood. Possible applications of this method range from bio-mechanical observations in fresh plant tissue to fracture mechanics aspects in structural timber.     

Transport and accumulation of PVP-Hypericin in cancer and normal cells characterized by image correlation spectroscopy techniques

PVP-Hypericin (PVP: polyvinylpyrrolidone) is a potent anti-cancer photosensitizer for photodynamic diagnosis (PDD) and therapy (PDT). However, cellular targets and mechanisms involved in the cancer-selectivity of the photosensitizer are not yet fully understood. This paper gives new insights into the differential transport and localization of PVP-Hypericin in cancer and normal cells which are essential to unravel the mechanisms of action and cancer-selectivity. Temporal (TICS) and spatiotemporal (STICS) image correlation spectroscopy are used for the assessment of PVP-Hypericin diffusion and/or velocity in the case of concerted flow in human cervical epithelial HeLa and human lung carcinoma A549 cells, as well as in human primary dendritic cells (DC) and human peripheral blood mononuclear cells (PBMC). Spatiotemporal image cross-correlation spectroscopy (STICCS) based on organelle specific fluorescent labeling is employed to study the accumulation of the photosensitizer in nucleus, mitochondria, early-endosomes and lysosomes of the cells and to assess the dynamics of co-migrating molecules. Whereas STICS and TICS did not show a remarkable difference between the dynamics of PVP-Hypericin in HeLa, A549 and DC cells, a significantly different diffusion rate of the photosensitizer was measured in PBMC. STICCS detected a stationary accumulation of PVP-Hypericin within the nucleus, mitochondria, early endosomes and lysosomes of HeLa and A549 cells. However, significant flow due to the directed motion of the organelles was detected. In contrast, no accumulation in the nucleus and mitochondria of DC and PBMC could be monitored.     

A video image correlation technique for the measurement of electrophoretic mobility

The electrophoretic movement of blood cells has been examined with a video image correlator using the signal from a 625 line monochrome television camera attached to the optical arrangement. The diffraction pattern of a cell produces a characteristic signal which can be detected and registered in any television scan line to form the data of the first image frame. Under the influence of an electric field, cells moved across the television field of view and after a short interval appeared on different scan lines. The detection and registering process was repeated, and a cross-correlation function between the first and second image frame was computed. This function was stored in an output buffer which was continuously updated as fresh functions were integrated with the existing contents. The mean electrophoretic mobility of a suspension of particles can be calculated from the peak position of the integrated correlation function. Using a triangle deconvolution procedure, the relative contributions of subpopulations lymphocytes prepared from peripheral blood samples were examined.     

Effect of round curvature of anterior implant-supported zirconia frameworks: finite element analysis and in vitro study using digital image correlation

Two groups of 4-unit zirconia frameworks were produced by CAD/CAM to simulate the restoration of an anterior edentulous gap supported by 2 implant-abutment assemblies. Group 1 comprised straight configuration frameworks and group 2 consisted of arched frameworks. Specimens were made with the same connector cross-section area and were cemented and submitted to static loads. Displacements were captured with two high-speed photographic cameras and analysed with video correlation system. Frameworks and the implant-abutment assembly were scanned and converted to 3DCAD objects by reverse engineering process. A specimen of each group was veneered and the corresponding 3D geometry was similarly obtained after scanning. Numerical models were created from the CAD objects and the FE analysis was performed on the zirconia frameworks and on the FPDs bi-layered with porcelain (veneered frameworks). Displacements were higher for the curved frameworks group, under any load. The predicted displacements correlated well with the experimental values of the two framework groups, but on the straight framework the experimental vertical displacements were superior to those predicted by the FEA. The results showed that the round curvature of zirconia anterior implant-supported FPDs plays a significant role on the deformation/stress of FPDs that cannot be neglected neither in testing nor in simulation and should be considered in the clinical setting.     

Studies of distribution,location and dynamic properties of EGFR on the cell surface measured by image correlation spectroscopy

In this work, we have studied the distribution and dynamic properties of Epidermal Growth Factor (EGF) receptors in the plasma membrane of fixed and live cells as well as the extent of co-localization of this transmembrane protein with proteins specific for three-membrane microdomains: membrane rafts, caveolae and clathrin-coated pits. This was achieved using a family of image-processing tools called image correlation spectroscopy (ICS), image cross-correlation spectroscopy (ICCS) and dynamic image correlation spectroscopy (DICS). Our results indicate that EGFR is diffusely distributed on the cell surface at 37°C and aggregates as the temperature is lowered to 4°C. This aggregation takes place within 15 min and is reversible. Changes in temperature also affect the diffusion of EGFR by two orders of magnitude. The dynamic properties of EGFR are similar to the dynamic properties of a GPI-anchored protein known to be present in membrane rafts, which motivated us to explore the extent of co-localization of EGFR with this membrane raft protein using ICCS. Our results indicate that more than half of the EGFR population is present in membrane rafts and smaller percentages are present in caveolae and clathrin-coated pits.