Spatial Assessment of Heterogeneous Tissue Natural Frequency Using Micro-Force Optical Coherence Elastography.

Analysis of corneal tissue natural frequency was recently proposed as a biomarker for corneal biomechanics and has been performed using high-resolution optical coherence tomography (OCT)-based elastography (OCE). However, it remains unknown whether natural frequency analysis can resolve local variations in tissue structure. We measured heterogeneous samples to evaluate the correspondence between natural frequency distributions and regional structural variations. Sub-micrometer sample oscillations were induced point-wise by microliter air pulses (60-85 Pa, 3 ms) and detected correspondingly at each point using a 1,300 nm spectral domain common path OCT system with 0.44 nm phase detection sensitivity. The resulting oscillation frequency features were analyzed via fast Fourier transform and natural frequency was characterized using a single degree of freedom (SDOF) model. Oscillation features at each measurement point showed a complex frequency response with multiple frequency components that corresponded with global structural features; while the variation of frequency magnitude at each location reflected the local sample features. Silicone blocks (255.1 ± 11.0 Hz and 249.0 ± 4.6 Hz) embedded in an agar base (355.6 ± 0.8 Hz and 361.3 ± 5.5 Hz) were clearly distinguishable by natural frequency. In a beef shank sample, central fat and connective tissues had lower natural frequencies (91.7 ± 58.2 Hz) than muscle tissue (left side: 252.6 ± 52.3 Hz; right side: 161.5 ± 35.8 Hz). As a first step, we have shown the possibility of natural frequency OCE methods to characterize global and local features of heterogeneous samples. This method can provide additional information on corneal properties, complementary to current clinical biomechanical assessments, and could become a useful tool for clinical detection of ocular disease and evaluation of medical or surgical treatment outcomes.

Adaptive optics wavefront correction using a damped transpose matrix of the influence function.

To assess the performance of adaptive optics and predict an optimal wavefront correction, we built a wavefront reconstructor with a damped transpose matrix of the influence function. Using an integral control strategy, we tested this reconstructor with four deformable mirrors in an experimental system, an adaptive optics scanning laser ophthalmoscope, and an adaptive optics near-confocal ophthalmoscope. Testing results proved that this reconstructor could ensure a stable and precise correction for wavefront aberration compared to a conventional optimal reconstructor formed by the inverse matrix of the influence function. This method may provide a helpful tool for testing, evaluating, and optimizing adaptive optics systems.

In vivo measurement of the lineal density of red blood cells in human retinal capillaries using high-speed adaptive optics ophthalmoscopy.

We present an automated method for measuring the lineal density of red blood cells (RBCs) in human retinal capillaries using adaptive optics near-confocal ophthalmoscopy (AONCO). The spatiotemporal traces of RBCs flowing in retinal capillaries were extracted from AONCO images, enhanced using the Gabor filter, the vesselness filter, and the Hough transform. A total of 43 capillary segments (each 50 µm long) were analyzed in 12 eyes of 12 subjects, and the measurement error of the automated method was 0.09 cell over 50 µm compared with results obtained by manual counting. Our method provides a tool for assessing RBC spatial distribution in retinal capillaries.

Clinical Corneal Optical Coherence Elastography Measurement Precision: Effect of Heartbeat and Respiration.

Purpose: Normal physiological movements (e.g., respiration and heartbeat) induce eye motions during clinical measurements of human corneal biomechanical properties using optical coherence elastography (OCE). We quantified the effects of respiratory and cardiac-induced eye motions on clinical corneal OCE measurement precision and repeatability. Methods: Corneal OCE was performed using low-force, micro-air-pulse tissue stimulation and high-resolution phase-sensitive optical coherence tomography (OCT) imaging. Axial surface displacements of the corneal apex were measured (M-mode) at a 70-kHz sampling rate and three different stimulation pressures (20-60 Pa). Simultaneously, the axial corneal position was tracked with structural OCT imaging, while the heartrate and respiration were monitored over a 90 second period. Results: Respiratory- and cardiac-induced eye motions have distinctly lower frequency (0.1-1 Hz) and much greater amplitude (up to ± 50 µm movements) than air-pulse-induced corneal tissue deformations (∼250 Hz, <1 µm). The corneal displacements induced during OCE measurements in vivo were -0.41 ± 0.06 µm (n = 22 measurements, coefficient of variation [CV]: 14.6%) and -0.44 ± 0.07 µm (n = 50 measurements, CV: 15.9%), respectively, from two human subjects at 40 Pa stimulation pressure. Observed variation in corneal tissue displacements were not associated with tissue stimulation magnitude, or the amplitude of physiologically induced axial eye motion. Conclusions: The microsecond timescale and submicron tissue displacements observed during corneal OCE measurements are separable from normal involuntary physiological movements, such as the oculocardiac pulse and respiratory movements. Translational Relevance: This work advances innovations in biomedical imaging and engineering for clinical diagnostic applications for soft-tissue biomechanical testing.

An imaging system integrating optical coherence tomography and interferometry for in vivo measurement of the thickness and dynamics of the tear film.

BACKGROUND: The outermost layer of the tear film consists of a thin lipid layer (LL). The lipid layer serves as a barrier against evaporation of the aqueous component of the tear film. The ability to simultaneously image both the lipid layer thickness and overall tear film thickness is novel, and will help further understandings of mechanisms of how the lipid layer assembles and interacts with the full tear film thickness. METHODS: We developed a system that combines simultaneous optical coherence tomography (OCT) and thickness dependent fringes (TDF) interferometry for in vivo imaging of the tear film. The OCT possesses an axial resolution of 1.38 µm in tear film, providing an accurate measurement of the thickness of the overall tear film. The TDF can detect a minimal change of approximately 15 nm in LL thickness. In addition, the spatial resolution of TDF images in x-y plane is 5 µm. RESULTS: The effect of instilling artificial tears on the PCTF and PLTF was examined. In both contact lens and non-contact lens wear, it could be observed from the OCT results that instillation of artificial tears increased the thickness of the overall tear film immediately, followed by a gradual reduction thereafter. These findings were consistent with other studies. However, unlike those previous reports, the thickness of the LL in this study was quantified simultaneously with the TDF subsystem. The results showed that bulking agents such as these artificial tears were not necessarily intended to increase the LL thickness. Immediately after instillation of artificial tears, the PCTF increased from 4.4 ± 0.97 to 20.3 ± 3.6 µm. The PCTF then decreased to 8.8 ± 2.1 µm at 4 min post-instillation. The thicknesses of the LL were 62.4 ± 14.5 nm, 48.7 ± 5.3 nm, and 55.2 ± 9.8 nm at pre-drop instillation, post-drop instillation, and 4-min post-drop instillation, respectively. CONCLUSIONS: In this work, we have described a novel imaging system that integrated OCT and TDF imaging techniques, which may facilitate the study of many physiological and clinical aspects of the tear film.

Noninvasive in vivo characterization of erythrocyte motion in human retinal capillaries using high-speed adaptive optics near-confocal imaging.

The flow of erythrocytes in parafoveal capillaries was imaged in the living human eye with an adaptive optics near-confocal ophthalmoscope at a frame rate of 800 Hz with a low coherence near-infrared (NIR) light source. Spatiotemporal traces of the erythrocyte movement were extracted from consecutive images. Erythrocyte velocity was measured using custom software based on the Radon transform. The impact of imaging speed on velocity measurement was estimated using images of frame rates of 200, 400, and 800 Hz. The NIR light allowed for long imaging periods without visually stimulating the retina and disturbing the natural rheological state. High speed near-confocal imaging enabled direct and accurate measurement of erythrocyte velocity, and revealed a distinctively cardiac-dependent pulsatile velocity waveform of the erythrocyte flow in retinal capillaries, disclosed the impact of the leukocytes on erythrocyte motion, and provided new metrics for precise assessment of erythrocyte movement. The approach may facilitate new investigations on the pathophysiology of retinal microcirculation with applications for ocular and systemic diseases.

High speed adaptive optics ophthalmoscopy with an anamorphic point spread function.

Retinal imaging working with a line scan mechanism and a line camera has the potential to image the eye with a near-confocal performance at the high frame rate, but this regime has difficulty to collect sufficient imaging light while adequately digitize the optical resolution in adaptive optics imaging. To meet this challenge, we have developed an adaptive optics line scan ophthalmoscope with an anamorphic point spread function. The instrument uses a high-speed line camera to acquire the retinal image and act as a confocal gate. Meanwhile, it employs a digital micro-mirror device to modulate the imaging light into a line of point sources illuminating the retina. The anamorphic mechanism ensures adequate digitization of the optical resolution and increases light collecting efficiency. We demonstrate imaging of the living human retina with cellular level resolution at a frame rate of 200 frames/second (FPS) with a digitization of 512 × 512 pixels over a field of view of 1.2° × 1.2°. We have assessed cone photoreceptor structure in images acquired at 100, 200, and 800 FPS in 2 normal human subjects, and confirmed that retinal images acquired at high speed rendered macular cone mosaic with improved measurement repeatability.

High-speed adaptive optics line scan confocal retinal imaging for human eye.

PURPOSE: Continuous and rapid eye movement causes significant intraframe distortion in adaptive optics high resolution retinal imaging. To minimize this artifact, we developed a high speed adaptive optics line scan confocal retinal imaging system. METHODS: A high speed line camera was employed to acquire retinal image and custom adaptive optics was developed to compensate the wave aberration of the human eye's optics. The spatial resolution and signal to noise ratio were assessed in model eye and in living human eye. The improvement of imaging fidelity was estimated by reduction of intra-frame distortion of retinal images acquired in the living human eyes with frame rates at 30 frames/second (FPS), 100 FPS, and 200 FPS. RESULTS: The device produced retinal image with cellular level resolution at 200 FPS with a digitization of 512×512 pixels/frame in the living human eye. Cone photoreceptors in the central fovea and rod photoreceptors near the fovea were resolved in three human subjects in normal chorioretinal health. Compared with retinal images acquired at 30 FPS, the intra-frame distortion in images taken at 200 FPS was reduced by 50.9% to 79.7%. CONCLUSIONS: We demonstrated the feasibility of acquiring high resolution retinal images in the living human eye at a speed that minimizes retinal motion artifact. This device may facilitate research involving subjects with nystagmus or unsteady fixation due to central vision loss.

Adaptive optics parallel near-confocal scanning ophthalmoscopy.

We present an adaptive optics parallel near-confocal scanning ophthalmoscope (AOPCSO) using a digital micromirror device (DMD). The imaging light is modulated to be a line of point sources by the DMD, illuminating the retina simultaneously. By using a high-speed line camera to acquire the image and using adaptive optics to compensate the ocular wave aberration, the AOPCSO can image the living human eye with cellular level resolution at the frame rate of 100 Hz. AOPCSO has been demonstrated with improved spatial resolution in imaging of the living human retina compared with adaptive optics line scan ophthalmoscopy.

Association between MMP-12-82A/G polymorphism and cancer risk: a meta-analysis.

BACKGROUND: Numerous studies have focused on the association between MMP-12-82A>G polymorphism and cancer risk, but produced inconsistent results. Therefore, we performed a meta-analysis of case-control study to evaluate the association of MMP-12-82A>G polymorphism and cancer risk. METHODS: A systematic literature search was conducted among PubMed, Web of Science, Science Direct, China National Knowledge Infrastructure (CNKI) and Wangfang databases updated on May 1st, 2015. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were used to evaluate the strength of association between this polymorphism and cancer risk. RESULTS: A total of seventeen case-control studies with 7,450 cases and 7,348 controls were identified and analyzed. Overall, there was no statistically significant association between MMP-12-82A>G polymorphism and increased risk of cancer under all genetic models. Subgroup analysis by ethnicity observed that there is no strong relationship between MMP-12-82A>G polymorphism and cancer risk among Asian and European populations. Furthermore, stratified analysis based on the source of control revealed no statistically significant association between MMP-12-82A>G polymorphism and cancer risk either in hospital-based or population-based studies. However, when we stratified analysis based on cancer type, significant association was found in ovarian cancer, but not in other types of cancer. CONCLUSION: This meta-analysis suggests that MMP-12-82A>G polymorphism is not significantly associated with overall cancer risk. However, MMP-12-82A>G polymorphism may increase the susceptibility to ovarian cancer.