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Neurulation is a highly synchronized biomechanical process leading to the formation of the brain and spinal cord, and its failure leads to neural tube defects (NTDs). Although we are rapidly learning the genetic mechanisms underlying NTDs, the biomechanical aspects are largely unknown. To understand the correlation between NTDs and tissue stiffness during neural tube closure (NTC), we imaged an NTD murine model using optical coherence tomography (OCT), Brillouin microscopy and confocal fluorescence microscopy. Here, we associate structural information from OCT with local stiffness from the Brillouin signal of embryos undergoing neurulation. The stiffness of neuroepithelial tissues in Mthfd1l null embryos was significantly lower than that of wild-type embryos. Additionally, exogenous formate supplementation improved tissue stiffness and gross embryonic morphology in nullizygous and heterozygous embryos. Our results demonstrate the significance of proper tissue stiffness in normal NTC and pave the way for future studies on the mechanobiology of normal and abnormal embryonic development.
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Tubo Neural , Neurulação , Tomografia de Coerência Óptica , Animais , Feminino , Camundongos , Fenômenos Biomecânicos , Embrião de Mamíferos/metabolismo , Formiato-Tetra-Hidrofolato Ligase/genética , Formiato-Tetra-Hidrofolato Ligase/metabolismo , Formiatos/metabolismo , Metilenotetra-Hidrofolato Desidrogenase (NADP)/genética , Metilenotetra-Hidrofolato Desidrogenase (NADP)/metabolismo , Camundongos Knockout , Microscopia Confocal , Mutação/genética , Tubo Neural/metabolismo , Defeitos do Tubo Neural/genética , Defeitos do Tubo Neural/metabolismo , Defeitos do Tubo Neural/patologia , Neurulação/genética , Tomografia de Coerência Óptica/métodosRESUMO
BACKGROUND: The brain and spinal cord formation is initiated in the earliest stages of mammalian pregnancy in a highly organized process known as neurulation. Environmental or genetic interferences can impair neurulation, resulting in clinically significant birth defects known collectively as neural tube defects. The Fuz gene encodes a subunit of the CPLANE complex, a macromolecular planar polarity effector required for ciliogenesis. Ablation of Fuz in mouse embryos results in exencephaly and spina bifida, including dysmorphic craniofacial structures due to defective cilia formation and impaired Sonic Hedgehog signaling. RESULTS: We demonstrate that knocking Fuz out during embryonic mouse development results in a hypoplastic hindbrain phenotype, displaying abnormal rhombomeres with reduced length and width. This phenotype is associated with persistent reduction of ventral neuroepithelial stiffness in a notochord adjacent area at the level of the rhombomere 5. The formation of cranial and paravertebral ganglia is also impaired in these embryos. CONCLUSIONS: This study reveals that hypoplastic hindbrain development, identified by abnormal rhombomere morphology and persistent loss of ventral neuroepithelial stiffness, precedes exencephaly in Fuz ablated murine mutants, indicating that the gene Fuz has a critical function sustaining normal neural tube development and neuronal differentiation.
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In this study, we introduce a multifocal acoustic radiation force source that combines an ultrasound transducer and a 3D-printed acoustic lens for application in reverberant optical coherence elastography (Rev-OCE). An array of plano-concave acoustic lenses, each with an 11.8 mm aperture diameter, were used to spatially distribute the acoustic energy generated by a 1â MHz planar ultrasound transducer, producing multiple focal spots on a target plane. These focal spots generate reverberant shear wave fields detected by the optical coherence tomography (OCT) system. The effectiveness of the multifocal Rev-OCE system in probing mechanical properties with high resolution is demonstrated in layered gelatin phantoms.
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To understand the dynamics of tissue stiffness during neural tube formation and closure in a murine model, we have developed a multimodal, coaligned imaging system combining optical coherence tomography (OCT) and Brillouin microscopy. Brillouin microscopy can map the longitudinal modulus of tissue but cannot provide structural images. Thus, it is limited for imaging dynamic processes such as neural tube formation and closure. To overcome this limitation, we have combined Brillouin microscopy and OCT in one coaligned instrument. OCT provided depth-resolved structural imaging with a micrometer-scale spatial resolution to guide stiffness mapping by Brillouin modality. 2D structural and Brillouin frequency shift maps were acquired of mouse embryos at gestational day (GD) 8.5, 9.5, and 10.5 with the multimodal system. The results demonstrate the capability of the system to obtain structural and stiffness information simultaneously.
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Microscopia , Tomografia de Coerência Óptica , Animais , Camundongos , Microscopia/métodos , Imagem Multimodal , Tubo Neural , Tomografia de Coerência Óptica/métodosRESUMO
In this work, we present an ultra-fast line-field optical coherence elastography system (LF-OCE) with an 11.5 MHz equivalent A-line rate. The system was composed of a line-field spectral domain optical coherence tomography system based on a supercontinuum light source, Michelson-type interferometer, and a high-speed 2D spectrometer. The system performed ultra-fast imaging of elastic waves in tissue-mimicking phantoms of various elasticities. The results corroborated well with mechanical testing. Following validation, LF-OCE measurements were made in in situ and in in vivo rabbit corneas under various conditions. The results show the capability of the system to rapidly image elastic waves in tissues.
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Técnicas de Imagem por Elasticidade , Animais , Córnea , Elasticidade , Imagens de Fantasmas , Coelhos , Tomografia de Coerência ÓpticaRESUMO
In optical coherence elastography (OCE), air-pulse stimulation has been widely used to produce propagation of mechanical waves for elastic characterization of tissues. In this paper, we propose the use of spatial deformation spreading (SDS) on the surface of samples produced by air-pulse stimulation for the OCE of transverse isotropic tissues. Experiments in isotropic tissue-mimicking phantoms and anisotropic chicken tibialis muscle were conducted using a spectral-domain optical coherence tomography system synchronized with a confocal air-pulse stimulation. SDS measurements were compared with wave speeds values calculated at different propagation angles. We found an approximately linear relationship between shear wave speed and SDS in isotropic phantoms, which was confirmed with predictions made by the numerical integration of a wave propagation model. Experimental measurements in chicken muscle show a good agreement between SDS and surface wave speed taken along and across the axis of symmetry of the tissues, also called degree of anisotropy. In summary, these results demonstrated the capabilities of SDS produced by the air-pulse technique in measuring the shear elastic anisotropy of transverse isotropic tissues.
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INTRODUCTION AND HYPOTHESIS: Quantitative characterization of the birth canal and critical structures before delivery may provide risk assessment for maternal birth injury. The objective of this study was to explore imaging capability of an antepartum tactile imaging (ATI) probe. METHODS: Twenty randomly selected women older than 21 years with completed 35th week of pregnancy and a premise of vaginal delivery were enrolled in the feasibility study. The biomechanical data were acquired using the ATI probe with a double-curved surface, shaped according to the fetal skull and equipped with 168 tactile sensors and an electromagnetic motion tracking sensor. Software package COMSOL Multiphysics was used for finite element modeling. Subjects were asked for assessment of pain and comfort levels experienced during the ATI examination. RESULTS: All 20 nulliparous women were successfully examined with the ATI. Mean age was 27.8 ± 4.1 years, BMI 30.7 ± 5.8, and week of pregnancy 38.8 ± 1.4. Biomechanical mapping with the ATI allowed real-time observation of the probe location, applied load to the vaginal walls, and a 3D tactile image composition. The nonlinear finite element model describing the stress-strain relationship of the pelvic tissue was developed and used for calculation of Young's modulus (E). Average perineal elastic modulus was 11.1 ± 4.3 kPa, levator ani 4.8 ± 2.4 kPa, and symphysis-perineum distance was 30.1 ± 6.9 mm. The pain assessment level for the ATI examination was 2.1 ± 0.8 (scale 1-4); the comfort level was 2.05 ± 0.69 (scale 1-3). CONCLUSIONS: The antepartum examination with the ATI probe allowed measurement of the tissue elasticity and anatomical distances. The pain level was low and the comfort level was comparable with manual palpation.
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Técnicas de Imagem por Elasticidade , Diafragma da Pelve , Adulto , Estudos de Viabilidade , Feminino , Humanos , Imageamento Tridimensional , Parto , Diafragma da Pelve/diagnóstico por imagem , Períneo/diagnóstico por imagem , Gravidez , Adulto JovemRESUMO
SIGNIFICANCE: A novel imaging technology, dynamic optical coherence elastography (OCE), was adapted for clinical noninvasive measurements of corneal biomechanics. PURPOSE: Determining corneal biomechanical properties is a long-standing challenge. Elasticity imaging methods have recently been developed and applied for clinical evaluation of soft tissues in cancer detection, atherosclerotic plaque evaluation, surgical guidance, and more. Here, we describe the use of dynamic OCE to characterize mechanical wave propagation in the human cornea in vivo, thus providing a method for clinical determination of corneal biomechanical properties. METHODS: High-resolution phase-sensitive optical coherence tomography imaging was combined with microliter air-pulse tissue stimulation to perform dynamic elasticity measurements in 18 eyes of nine participants. Low-pressure (0.1 mmHg), spatiotemporally discreet (150 µm, 800 µs) tissue stimulation produced submicron-scale tissue deformations that were measured at multiple positions over a 1-mm2 area. Surface wave velocity was measured and used to determine tissue stiffness. Elastic wave propagation velocity was measured and evaluated as a function of IOP and central corneal thickness. RESULTS: Submicron corneal surface displacement amplitude (range, 0.005 to 0.5 µm) responses were measured with high sensitivity (0.24 nm). Corneal elastic wave velocity ranged from 2.4 to 4.2 m/s (mean, 3.5; 95% confidence interval, 3.2 to 3.8 m/s) and was correlated with central corneal thickness (r = 0.64, P < .001) and IOP (r = 0.52, P = .02). CONCLUSIONS: Phase-sensitive optical coherence tomography imaging combined with microliter air-pulse mechanical tissue stimulation has sufficient detection sensitivity to observe submicron elastic wave propagation in corneal tissue. These measurements enable in vivo corneal stiffness determinations that will be further studied for use with disease detection and for monitoring clinical interventions.
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Córnea/diagnóstico por imagem , Técnicas de Imagem por Elasticidade/métodos , Elasticidade/fisiologia , Tomografia de Coerência Óptica/métodos , Adulto , Fenômenos Biomecânicos/fisiologia , Córnea/fisiologia , Feminino , Humanos , Masculino , Projetos PilotoRESUMO
We present an air-coupled ultrasonic radiation force probe co-focused with a phase-sensitive optical coherence tomography (OCT) system for quantitative wave-based elastography. A custom-made 1 MHz spherically focused piezoelectric transducer with a concentric 10 mm wide circular opening allowed for confocal micro-excitation of waves and phase-sensitive OCT imaging. Phantom studies demonstrated the capabilities of this probe to produce quasi-harmonic excitation up to 4 kHz for generation of elastic waves. Experimental results in ocular tissues showed highly detailed 2D and 3D elasticity mapping using this approach with great potential for clinical translation.
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Ar , Fenômenos Mecânicos , Tomografia de Coerência Óptica/métodos , Ondas Ultrassônicas , Fenômenos BiomecânicosRESUMO
We present an erratum to correct an inadvertent error made during the calculations of the in-focus fluence of pulsed laser used to excite nanoparticles [Opt. Lett.44, 3162 (2019)OPLEDP0146-959210.1364/OL.44.003162] and to update the conclusion regarding laser safety limits achieved with this type of excitation.
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Wave-based optical coherence elastography (OCE) is a rapidly emerging technique for elasticity assessment of tissues having high displacement sensitivity and simple implementation. However, most current noncontact wave excitation techniques are unable to target a specific tissue site in 3D and rely on transversal scanning of the imaging beam. Here, we demonstrate that dye-loaded perfluorocarbon nanoparticles (nanobombs) excited by a pulsed laser can produce localized axially propagating longitudinal shear waves while adhering to the laser safety limit. A phase-correction method was developed and implemented to perform sensitive nanobomb elastography using a â¼1.5 MHz Fourier domain mode-locking laser. The nanobomb activation was also monitored by detecting photoacoustic signals. The highly localized elastic waves detected by the nanobomb OCE suggest the possibility of high-resolution 3D elastographic imaging.
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Wave-based optical elastography is rapidly emerging as a powerful technique for quantifying tissue biomechanical properties due to its noninvasive nature and high displacement sensitivity. However, current approaches are limited in their ability to produce high-frequency waves and highly localized mechanical stress. In this Letter, we demonstrate that the rapid liquid-to-gas phase transition of dye-loaded perfluorocarbon nanodroplets ("nanobombs") initiated by a pulsed laser can produce highly localized, high-frequency, and broadband elastic waves. The waves were detected by an ultra-fast line-field low-coherence holography system. For comparison, we also excited waves using a focused micro-air-pulse. Results from tissue-mimicking phantoms showed that the nanobombs produced elastic waves with frequencies up to â¼9 kHz, which was much greater than the â¼2 kHz waves excited by the air-pulse. Consequently, the nanobombs enabled more accurate quantification of sample viscoelasticity. Combined with their potential for functionalization, the nanobombs show promise for accurate and highly specific noncontact all-optical elastography.
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Carbocianinas/química , Módulo de Elasticidade , Técnicas de Imagem por Elasticidade/instrumentação , Fluorocarbonos/química , Imagens de Fantasmas , Tomografia de Coerência Óptica/instrumentação , Fenômenos Biomecânicos , Técnicas de Imagem por Elasticidade/métodos , Lasers de Estado Sólido , Microesferas , Estresse Mecânico , Tomografia de Coerência Óptica/métodosRESUMO
SIGNIFICANCE: Measured corneal biomechanical properties are driven by intraocular pressure, tissue thickness, and inherent material properties. We demonstrate tissue thickness as an important factor in the measurement of corneal biomechanics that can confound short-term effects due to UV riboflavin cross-linking (CXL) treatment. PURPOSE: We isolate the effects of tissue thickness on the measured corneal biomechanical properties using optical coherence elastography by experimentally altering the tissue hydration state and stiffness. METHODS: Dynamic optical coherence elastography was performed using phase-sensitive optical coherence tomography imaging to quantify the tissue deformation dynamics resulting from a spatially discrete, low-force air pulse (150-µm spot size; 0.8-millisecond duration; <10 Pa [<0.08 mmHg]). The time-dependent surface deformation is characterized by a viscoelastic tissue recovery response, quantified by an exponential decay constant-relaxation rate. Ex vivo rabbit globes (n = 10) with fixed intraocular pressure (15 mmHg) were topically instilled every 5 minutes with 0.9% saline for 60 minutes and 20% dextran for another 60 minutes. Measurements were made after every 20 minutes to determine the central corneal thickness (CCT) and the relaxation rates. Cross-linking treatment was performed on another 13 eyes, applying isotonic riboflavin (n = 6) and hypertonic riboflavin (n = 7) every 5 minutes for 30 minutes, followed by UV irradiation (365 nm, 3 mW/cm) for 30 minutes while instilling riboflavin. Central corneal thickness and relaxation rates were obtained before and after CXL treatment. RESULTS: Corneal thickness was positively correlated (R = 0.9) with relaxation rates. In the CXL-treated eyes, isotonic riboflavin did not affect CCT and showed a significant increase in relaxation rates (+10%; P = .01) from 2.29 ms to 2.53 ms. Hypertonic riboflavin showed a significant CCT decrease (-31%; P = .01) from 618 µm to 429 µm but showed little change in relaxation rates after CXL treatment. CONCLUSIONS: Corneal thickness and stiffness are correlated positively. A higher relaxation rate implied stiffer material properties after isotonic CXL treatment. Hypertonic CXL treatment results in a stiffness decrease that offsets the stiffness increase with CXL treatment.
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Córnea/fisiopatologia , Técnicas de Imagem por Elasticidade , Elasticidade/fisiologia , Fármacos Fotossensibilizantes/farmacologia , Riboflavina/farmacologia , Animais , Fenômenos Biomecânicos , Colágeno/metabolismo , Córnea/diagnóstico por imagem , Substância Própria/efeitos dos fármacos , Substância Própria/metabolismo , Reagentes de Ligações Cruzadas , Pressão Intraocular/efeitos dos fármacos , Tamanho do Órgão , Concentração Osmolar , Coelhos , Tomografia de Coerência Óptica , Tonometria Ocular , Raios UltravioletaRESUMO
Contrast-enhanced ultrasound imaging utilizing the nonlinear responses of microbubbles is proposed for identifying ventricular shunt malfunction. The developed method suppresses the signal from walls of a shunt catheter and background tissues, and allows accurate measurements of the cerebrospinal fluid flow within the shunt catheter using a relatively small concentration of microbubbles. The flow rates estimated in the linear mode were significantly underestimated (65% at 0.1 ml/min and 5.0% microbubble concentration) while estimates using the nonlinear mode were not. Overall, the nonlinear responses of microbubbles improve the estimation of flow rates in a shunt catheter at low concentrations of microbubbles.
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The mechanical properties of tissues can provide valuable information about tissue integrity and health and can assist in detecting and monitoring the progression of diseases such as keratoconus. Optical coherence elastography (OCE) is a rapidly emerging technique, which can assess localized mechanical contrast in tissues with micrometer spatial resolution. In this work we present a noncontact method of optical coherence elastography to evaluate the changes in the mechanical properties of the cornea after UV-induced collagen cross-linking. A focused air-pulse induced a low amplitude (µm scale) elastic wave, which then propagated radially and was imaged in three dimensions by a phase-stabilized swept source optical coherence tomography (PhS-SSOCT) system. The elastic wave velocity was translated to Young's modulus in agar phantoms of various concentrations. Additionally, the speed of the elastic wave significantly changed in porcine cornea before and after UV-induced corneal collagen cross-linking (CXL). Moreover, different layers of the cornea, such as the anterior stroma, posterior stroma, and inner region, could be discerned from the phase velocities of the elastic wave. Therefore, because of noncontact excitation and imaging, this method may be useful for in vivo detection of ocular diseases such as keratoconus and evaluation of therapeutic interventions such as CXL.
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Significance: The biomechanical impact of refractive surgery has long been an area of investigation. Changes to the cornea structure cause alterations to its mechanical integrity, but few studies have examined its specific mechanical impact. Aim: To quantify how the biomechanical properties of the cornea are altered by laser assisted in situ keratomileusis (LASIK) using optical coherence elastography (OCE) in ex vivo porcine corneas. Approach: Three OCE techniques, wave-based air-coupled ultrasound (ACUS) OCE, heartbeat (Hb) OCE, and compression OCE were used to measure the mechanical properties of paired porcine corneas, where one eye of the pair was left untreated, and the fellow eye underwent LASIK. Changes in stiffness as a function of intraocular pressure (IOP) before and after LASIK were measured using each technique. Results: ACUS-OCE showed that corneal stiffness changed as a function of IOP for both the untreated and the treated groups. The elastic wave speed after LASIK was lower than before LASIK. Hb-OCE and compression OCE showed regional changes in corneal strain after LASIK, where the absolute strain difference between the cornea anterior and posterior increased after LASIK. Conclusions: The results of this study suggest that LASIK may soften the cornea and that these changes are largely localized to the region where the surgery was performed.
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Técnicas de Imagem por Elasticidade , Oftalmopatias , Ceratomileuse Assistida por Excimer Laser In Situ , Animais , Suínos , Ceratomileuse Assistida por Excimer Laser In Situ/métodos , Fenômenos Biomecânicos , Córnea/diagnóstico por imagem , Córnea/cirurgia , Tonometria OcularRESUMO
Acute alcohol ingestion has been found to impact visual functions, including eye movement, but its effects on corneal biomechanical properties remain unclear. This study aimed to investigate the influence of acute alcohol consumption on corneal biomechanical properties using optical coherence elastography (OCE). An air-coupled ultrasound transducer induced elastic waves in mice corneas in vivo, and a high-resolution phase-sensitive optical coherence tomography (OCT) system tracked the mechanical waves to quantify the elastic wave speed. In vivo measurements were performed on three groups of age- and gender-matched mice: control, placebo (administered saline), and alcohol (administered ethanol) groups. Longitudinal measurements were conducted over a one-hour period to assess acute temporal changes in wave speeds, which are associated with inherent biomechanical properties of the cornea. The results showed a significant decrease in wave speed for the alcohol group after 10 min of ingestion in comparison to pre-ingestion values (p = 0.0096), whereas the temporal wave speed changes for the placebo group were statistically insignificant (p = 0.076). In contrast, the control group showed no significant changes in elastic wave speed and corneal thickness. Furthermore, a significant difference was observed between the wave speeds of the placebo and alcohol groups at each measurement time point between 10 and 50 min (p < 0.05), though both groups exhibited a similar trend in corneal thickness change. The findings of this study have important implications for clinical assessments and research in corneal disorders, highlighting the potential of OCE as a valuable tool for evaluating such changes.
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Consumo de Bebidas Alcoólicas , Córnea , Técnicas de Imagem por Elasticidade , Córnea/diagnóstico por imagem , Córnea/fisiologia , Animais , Técnicas de Imagem por Elasticidade/métodos , Camundongos , Consumo de Bebidas Alcoólicas/fisiopatologia , Fenômenos Biomecânicos , Masculino , Tomografia de Coerência Óptica/métodos , Etanol , Camundongos Endogâmicos C57BL , Elasticidade , FemininoRESUMO
During the first month of pregnancy, the brain and spinal cord are formed through a process called neurulation. However, this process can be altered by low serum levels of folic acid, environmental factors, or genetic predispositions. In 2018, a surveillance study in Botswana, a country with a high incidence of human immunodeficiency virus (HIV) and lacking mandatory food folate fortification programs, found that newborns whose mothers were taking dolutegravir (DTG) during the first trimester of pregnancy had an increased risk of neural tube defects (NTDs). As a result, the World Health Organization and the U.S. Food and Drug Administration have issued guidelines emphasizing the potential risks associated with the use of DTG-based antiretroviral therapies during pregnancy. To elucidate the potential mechanisms underlying the DTG-induced NTDs, we sought to assess the potential neurotoxicity of DTG in stem cell-derived brain organoids. The gene expression of brain organoids developed in the presence of DTG was analyzed by RNA sequencing, Optical Coherence Tomography (OCT), Optical Coherence Elastography (OCE), and Brillouin microscopy. The sequencing data shows that DTG induces the expression of the folate receptor (FOLR1) and modifies the expression of genes required for neurogenesis. The Brillouin frequency shift observed at the surface of DTG-exposed brain organoids indicates an increase in superficial tissue stiffness. In contrast, reverberant OCE measurements indicate decreased organoid volumes and internal stiffness.
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The crystalline lens is a transparent, biconvex structure that has its curvature and refractive power modulated to focus light onto the retina. This intrinsic morphological adjustment of the lens to fulfill changing visual demands is achieved by the coordinated interaction between the lens and its suspension system, which includes the lens capsule. Thus, characterizing the influence of the lens capsule on the whole lens's biomechanical properties is important for understanding the physiological process of accommodation and early diagnosis and treatment of lenticular diseases. In this study, we assessed the viscoelastic properties of the lens using phase-sensitive optical coherence elastography (PhS-OCE) coupled with acoustic radiation force (ARF) excitation. The elastic wave propagation induced by ARF excitation, which was focused on the surface of the lens, was tracked with phase-sensitive optical coherence tomography. Experiments were conducted on eight freshly excised porcine lenses before and after the capsular bag was dissected away. Results showed that the group velocity of the surface elastic wave, V , in the lens with the capsule intact ( V = 2.55 ± 0.23 m / s ) was significantly higher (p < 0.001) than after the capsule was removed ( V = 1.19 ± 0.25 m / s ). Similarly, the viscoelastic assessment using a model that utilizes the dispersion of a surface wave showed that both Young's modulus, E, and shear viscosity coefficient, η, of the encapsulated lens ( E = 8.14 ± 1.10 k P a , η = 0.89 ± 0.093 P a â s ) were significantly higher than that of the decapsulated lens ( E = 3.10 ± 0.43 k P a , η = 0.28 ± 0.021 P a â s ). These findings, together with the geometrical change upon removal of the capsule, indicate that the capsule plays a critical role in determining the viscoelastic properties of the crystalline lens.
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Significance: Quantifying the biomechanical properties of the whole eye globe can provide a comprehensive understanding of the interactions among interconnected ocular components during dynamic physiological processes. By doing so, clinicians and researchers can gain valuable insights into the mechanisms underlying ocular diseases, such as glaucoma, and design interventions tailored to each patient's unique needs. Aim: The aim of this study was to evaluate the feasibility and effectiveness of a multifocal acoustic radiation force (ARF) based reverberant optical coherence elastography (RevOCE) technique for quantifying shear wave speeds in different ocular components simultaneously. Approach: We implemented a multifocal ARF technique to generate reverberant shear wave fields, which were then detected using phase-sensitive optical coherence tomography. A 3D-printed acoustic lens array was employed to manipulate a collimated ARF beam generated by an ultrasound transducer, producing multiple focused ARF beams on mouse eye globes ex vivo. RevOCE measurements were conducted using an excitation pulse train consisting of 10 cycles at 3 kHz, followed by data processing to produce a volumetric map of the shear wave speed. Results: The results show that the system can successfully generate reverberant shear wave fields in the eye globe, allowing for simultaneous estimation of shear wave speeds in various ocular components, including cornea, iris, lens, sclera, and retina. A comparative analysis revealed notable differences in wave speeds between different parts of the eye, for example, between the apical region of the cornea and the pupillary zone of the iris (p=0.003). Moreover, the study also revealed regional variations in the biomechanical properties of ocular components as evidenced by greater wave speeds near the apex of the cornea compared to its periphery. Conclusions: The study demonstrated the effectiveness of RevOCE based on a non-invasive multifocal ARF for assessing the biomechanical properties of the whole eyeball. The findings indicate the potential to provide a comprehensive understanding of the mechanical behavior of the whole eye, which could lead to improved diagnosis and treatment of ocular diseases.