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Significance: Three-dimensional photoacoustic imaging (PAM) has emerged as a promising technique for non-invasive label-free visualization and characterization of biological tissues with high spatial resolution and functional contrast., Aim: The application of PAM and ultrasound as a microscopy technique of study for Atlantic salmon skin is presented here., Approach: A custom ultrasound and photoacoustic experimental setup was used for conducting this experiment with a sample preparation method where the salmon skin is embedded in agarose and lifted from the bottom of the petridish., Results: The results of C-scan, B-scan, and overlayed images of ultrasound and photoacoustic are presented. The results are then analyzed for understanding the pigment map and its relation to salmon behavior to external stimuli. The photoacoustic images are compared with the optical images and analyzed further. A custom colormap and alpha map is designed and the matrices responsible for PAM and ultrasound are inserted together to overlay the ultrasound image and PAM image on top of each other., Conclusions: In this study, we propose an approach that combines scanning acoustic microscopy (SAM) images with PAM images for providing a comprehensive understanding of the salmon skin tissue. Overlaying acoustic and photoacoustic images enabled unique visualization of tissue morphology, with respect to identification of structural features in the context of their pigment distribution., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ranjan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Purpose: Phakic lens implantation in the ciliary sulcus of the eye can be complicated by coincident ciliary body cysts (CBC). We developed an ultrasound imaging and mapping protocol for these cysts., Methods: This is a retrospective case series of all patients who underwent ICL workup at a single institution from April 2015 to October 2019. A standardized ultrasound biomicroscopy (UBM) imaging protocol was developed to screen for CBCs in either the ciliary body or sulcus. The locations and dimensions of all CBCs were graphically represented., Results: The prevalence of CBCs in 158 patients undergoing ICL workup was 34.8%. Among the 159 CBCs detected in 55 patients, 83 were in the sulcus (52%) and 76 were restricted to the ciliary body (48%). ICLs were implanted in 40 eyes with CBCs and 3 eyes with CBCs located within the sulcus horizontally required ICL repositioning due to ICL rotation or iris chafing., Conclusion: CBCs were incidentally found in 34.8% of patients undergoing ICL workup. ICL implantation was complicated in 3 of the eyes with CBCs in the horizontal sulcus. Although CBCs are not an absolute contraindication for ICL surgery, we recommend preoperative UBM screening of the ciliary sulcus., (© 2024. The Author(s).)

In Ultrasound Localization Microscopy (ULM), achieving high-resolution images relies on the precise localization of contrast agent particles across a series of beamformed frames. However, our study uncovers an enormous potential: The process of delay-and-sum beamforming leads to an irreversible reduction of Radio-Frequency (RF) channel data, while its implications for localization remain largely unexplored. The rich contextual information embedded within RF wavefronts, including their hyperbolic shape and phase, offers great promise for guiding Deep Neural Networks (DNNs) in challenging localization scenarios. To fully exploit this data, we propose to directly localize scatterers in RF channel data. Our approach involves a custom super-resolution DNN using learned feature channel shuffling, non-maximum suppression, and a semi-global convolutional block for reliable and accurate wavefront localization. Additionally, we introduce a geometric point transformation that facilitates seamless mapping to the B-mode coordinate space. To understand the impact of beamforming on ULM, we validate the effectiveness of our method by conducting an extensive comparison with State-Of-The-Art (SOTA) techniques. We present the inaugural in vivo results from a wavefront-localizing DNN, highlighting its real-world practicality. Our findings show that RF-ULM bridges the domain shift between synthetic and real datasets, offering a considerable advantage in terms of precision and complexity. To enable the broader research community to benefit from our findings, our code and the associated SOTA methods are made available at https://github.com/hahnec/rf-ulm.

Objective: To compare anterior chamber depth (ACD) and lens thickness (LT) measurements by ultrasound biomicroscopy (UBM), A-scan cross vector (CV) overlay with UBM, and immersion A-scan technique in pediatric eyes., Methods: This prospective comparative cohort study comprised 43 eyes of 25 pediatric participants (mean age: 2.3±2.2 y). UBM and immersion A-scan biometry were performed prior to dilation and intraocular surgery. ACD and LT were measured by UBM image analysis, A-scan CV UBM overlay, and immersion A-scan technique., Results: ACD and LT measurements obtained using immersion A-scan were significantly greater than with UBM image analysis with mean differences of 0.52 mm and 0.62 mm, respectively (p < 0.001). Immersion A-scan and UBM measurements were moderately correlated (r = 0.70 and 0.64, p < 0.001). ACD and LT measurements obtained using CV overlay were not significantly different than UBM measurements and the values were strongly positively correlated (r = 0.95 and 0.93, p < 0.001)., Conclusion: Immersion A-scan may overestimate ACD and LT compared to UBM in pediatric patients due to oblique placement of the A-scan probe relative to the optical axis. Supplemental use of UBM and/or CV overlay is indicated to improve measurement accuracy in pediatric patients who cannot reliably fixate due to the ability to confirm proper alignment of the probe with the pupil by visualizing the anterior segment., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2024 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Objective: This work aimed to promote the interaction of a modified gas vesicle (GV) with cathepsin B (CTSB) protease and analysed their backscattered signal by ultrasound (US)., Methods: We modified the sequence of the gene coding for GvpC to contain a CTSB cleavage and expressed the protein in an Escherichia coli recombinant system. The protein was purified and added to GVs preparations in which the original GvpC was removed (ΔGV), constituting the modified GV (GV*). Western blot testing was used to compare GVs with GvpC and engineered GvpC at starting (T0) and after 24 h (T24) reacting with CTSB. A 21 MHz US B-mode and non-linear contrast mode (5% total power) imaged US phantoms having samples of GVwt, ΔGV (stripped GV), GV* and CTSB + GV*. Also, a 21 MHz US B-mode imaged US phantoms having a tumour cell line extracellular fraction (TCEF) and the TCEF + GV* sample. A 100% total US power was applied to collapse the GV structure., Results: On Western blotting, we detected a decrease in engineered GvpC levels 24 h after the incubation of GV* with CTSB, compared with the concentration at T0, suggesting that CTSB cleaved the engineered GvpC. Regions-of-interest over image of phantom cross-sections were determined and the B-mode image mean grey-level intensity resulted in a significant (p < 0.05) increase comparing CTSB + GV* with PBS (control), GVwt, ΔGV and GV*. Non-linear mode image grey-level intensity from CTSB + GV* increased by 11.79, 7.86 and 14.75 dB from samples containing GVwt, ΔGV and GV*, respectively. GV preparations incubated with TCEF and the TCEF + GV* sample showed an increase of 81% in signal compared with TCEF + GVwt., Conclusion: The increased US backscattered signal intensity suggests GVs as a potential biosensor for protease activity, possibly aiding the detection of protease-rich tissue regions., Competing Interests: Conflict of interest The authors declare that they have no conflict of interest., (Copyright © 2024 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Ultrasound Localization Microscopy (ULM) facilitates structural and hemodynamic imaging of microvessels with a resolution of tens of micrometers. In ULM, the extraction of effective microbubble signals is crucial for image quality. Singular Value Decomposition (SVD) is currently the most prevalent method for microbubble signal extraction in ULM. Most existing ULM studies employ a fixed SVD filter threshold using empirical values which will lead to imaging quality degradation due to the insufficient separation of blood signals. In this study, we propose an adaptive and non-threshold SVD filter based on canopy-density clustering, termed DCC-SVD. This filter automatically classifies the components of the SVD based on the density of their spatiotemporal features, eliminating the need for parameter selection. In in vitro tube phantom, DCC-SVD demonstrated its ability to adaptive separation of blood and bubble signal at varying microbubble concentrations and flow rates. We compared the proposed DCC-SVD method with the Block-match 3D (BM3D) filter and a classical adaptive method called spatial similarity matrix (SSM), using concentration-variable in vivo rat brain data, as well as open-source rat kidney and mouse tumor datasets. The proposed DCC-SVD improved the global spatial resolution by approximately 4 μm from 30.39 μm to 26.02 μm. It also captured vessel structure absent in images obtained by other methods and yielded a smoother vessel intensity profile, making it a promising spatiotemporal filter for ULM imaging., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Accurate prediction of postoperative vault in implantable collamer lens (ICL) implantation is crucial; however, current formulas often fail to account for individual anatomical variations, leading to suboptimal visual outcomes and necessitating improved predictive models. We aimed to verify the prediction accuracy of our new predictive model for vaulting based on anterior and posterior chamber structural parameters., Methods: This retrospective observational study included 137 patients (240 eyes) who previously underwent ICL surgery. Patients were randomly divided into the model establishment (192 eyes) or validation (48 eyes) groups. Preoperative measurements of the anterior and posterior chamber structures were obtained using Pentacam, CASIA2 anterior segment optical coherence tomography (AS-OCT), ultrasound biomicroscopy, and other devices. Stepwise multiple linear regression analysis was used to evaluate the relationship between the vault and each variable (WL formula). The Friedman test was performed for the vaulting prediction results of the WL, NK (Ver. 3), and KS formulas (Ver. 4) in CASIA2 AS-OCT, as well as the Zhu formula and vault measurements. The proportions of prediction error within ± 250 μm per formula were compared., Results: The predicted vault values of the WL, NK, KS, and Zhu formulas and vault measurements were 668.74 ± 162.12, 650.85 ± 248.47, 546.56 ± 128.99, 486.56 ± 210.76, and 716.06 ± 233.84 μm, respectively, with a significant difference (χ 2  = 69.883, P = 0.000). Significant differences were also found between the measured vault value and Zhu formula, measured vault value and KS formula, WL formula and Zhu formula, WL formula and KS formula, NK formula and KS formula, and NK formula and Zhu formula (P < 0.001) but not between other groups. The proportions of prediction error within ± 250 μm per formula were as follows: WL formula (81.3%) > NK formula (70.8%) > KS formula (66.7%) > Zhu formula (54.2%)., Conclusions: The WL formula, which considers the complexity of the anterior and posterior chamber structures, demonstrates greater calculation accuracy, compared with the KS (Ver. 4) and Zhu formulas. The proportion of absolute prediction error ≤ 250 μm is higher with the WL formula than with the NK formula (ver. 3). This enhanced predictive capability can improve ICL sizing decisions, thereby increasing the safety and efficacy of ICL implantation surgeries., (© 2024. The Author(s).)

Purpose: To determine the clinical outcomes in patients after type 1 Boston keratoprosthesis surgery and the significance of ultrasound biomicroscopy imaging for postoperative follow-up., Methods: This retrospective analysis included 20 eyes of 19 patients who underwent corneal transplantation with type 1 Boston keratoprosthesis between April 2014 and December 2021. Data on patient demographics, preoperative diagnosis, visual acuity, and postoperative clinical findings were analyzed., Results: Type 1 Boston keratoprosthesis implantation resulted in intermediate- and long-term positive outcomes. However, blindness and other serious complications such as glaucoma, retroprosthetic membrane formation, endophthalmitis, or retinal detachment also occurred. The use of ultrasound biomicroscopy imaging allowed for better evaluation of the back of the titanium plate, anterior segment structures, and the relationship of the prosthesis with surrounding tissues, which provided valuable postoperative information., Conclusion: Regular lifetime monitoring and treatment are necessary in patients who undergo Boston type 1 keratoprosthesis implantation for high-risk corneal transplantation. ultrasound biomicroscopy imaging can be a valuable imaging technique for the evaluation of patients with Boston type 1 keratoprosthesis, providing important information on anterior segment anatomy and potential complications. Further studies and consensus on postoperative follow-up protocols are required to optimize the management of patients with Boston type 1 keratoprosthesis.

Objective: This study aimed to develop and evaluate a deep learning-based model that could automatically measure anterior segment (AS) parameters on preoperative ultrasound biomicroscopy (UBM) images of implantable Collamer lens (ICL) surgery candidates., Methods: A total of 1164 panoramic UBM images were preoperatively obtained from 321 patients who received ICL surgery in the Eye Center of Renmin Hospital of Wuhan University (Wuhan, China) to develop an imaging database. First, the UNet++ network was utilized to segment AS tissues automatically, such as corneal lens and iris. In addition, image processing techniques and geometric localization algorithms were developed to automatically identify the anatomical landmarks (ALs) of pupil diameter (PD), anterior chamber depth (ACD), angle-to-angle distance (ATA), and sulcus-to-sulcus distance (STS). Based on the results of the latter two processes, PD, ACD, ATA, and STS can be measured. Meanwhile, an external dataset of 294 images from Huangshi Aier Eye Hospital was employed to further assess the model's performance in other center. Lastly, a subset of 100 random images from the external test set was chosen to compare the performance of the model with senior experts., Results: Whether in the internal test dataset or external test dataset, using manual labeling as the reference standard, the models achieved a mean Dice coefficient exceeding 0.880. Additionally, the intra-class correlation coefficients (ICCs) of ALs' coordinates were all greater than 0.947, and the percentage of Euclidean distance distribution of ALs within 250 μm was over 95.24%.While the ICCs for PD, ACD, ATA, and STS were greater than 0.957, furthermore, the average relative error (ARE) of PD, ACD, ATA, and STS were below 2.41%. In terms of human versus machine performance, the ICCs between the measurements performed by the model and those by senior experts were all greater than 0.931., Conclusion: A deep learning-based model could measure AS parameters using UBM images of ICL candidates, and exhibited a performance similar to that of a senior ophthalmologist., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2024. Published by Elsevier Inc.)

Purpose: To define the regular upper eyelid anatomy and their changes during dynamic motion using ultrasonographically depicted structures., Design: A cross-sectional study., Methods: High-resolution ultrasound biomicroscopy (50MHz) was performed on 84 upper eyelids of 42 healthy volunteers without evidence of eyelid conditions. The skin-orbicularis oculi complex (SOOC), levator aponeurosis, and Müller's muscle-conjunctival complex were imaged. The thickness of these structures was measured in primary gaze and downgaze positions., Results: SOOC and levator aponeurosis thickness was bigger with eyes open than with eyes closed (p < 0.01). With eyes closed, the thickness of SOOC on the pupillary midline and levator aponeurosis was positively correlated with age (p < 0.05, p < 0.01, respectively), and the thickness of levator aponeurosis and SOOC on the pupil midline was also positively correlated with BMI index (p < 0.05). Similarly, SOOC thickness on the medial and lateral canthus line is also positively correlated with BMI index and age (p < 0.05, p < 0.01, respectively). As for the differences between genders, the mean thickness of SOOC and levator aponeurosis was statistically different (p < 0.05, respectively). The magnitude of levator aponeurosis thickness in men varied more than in women in different ocular positions (p < 0.05). All measurements had an intramethod between 0.624 and 0.792, and inter-rater ICC and intrarater ICC between 0.748 and 0.850., Conclusions: Ultrasound biomicroscopy represents a noninvasive tool for the visualization of upper eyelid morphology. Expanding its application can help to understand the dynamics of upper eyelid physiological movement, aging, and disease research in different populations and evaluate surgical outcomes., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2024. Published by Elsevier Inc.)

Introduction: Intra-ocular lens (IOL) calcification is a rare yet serious complication, presenting as clouding within the optics of IOL and necessitating IOL exchange., Patient and Clinical Findings: In this case, a 77-year-old male experienced gradual vision loss in his left eye (LE) over four years post-cataract surgery a decade before. His best corrected visual acuity (BCVA) was 3/60 in the LE, with the anterior segment displaying a clear cornea but an opacified IOL within the capsular bag. Ultrasound biomicroscopy (UBM) revealed hyperechoic reflections from the IOL in the LE. ImageJ software used on UBM images indicated a significant difference in optical density (OD) compared to the right eye (RE)., Results: The patient was diagnosed with IOL calcification in LE, and underwent anterior vitrectomy and IOL exchange with sulcus-placed IOL with optic capture in the capsular bag, resulting in an improved visual acuity of 6/18. Microscopic examination of explanted IOL revealed whitish calcium carbonate crystal deposits on the IOL, which were visible under polarized light., Conclusions and Significance: The innovative use of polarized light and ImageJ software can be used in resource-constraint settings to confirm the diagnosis in such cases. Based on ImageJ on the UBM image, OD can pick up even subtle changes in the optical clarity of the IOL optic preoperatively., Competing Interests: The authors state no conflict of interest., (© 2024 The Authors.)

Objectives: Silicone oil (SO) is a commonly used intraocular tamponade in the treatment of rhegmatogenous retinal detachment (RRD). SO emulsification is a common complication after SO injection. This study aimed to investigate correlations between SO emulsification signs on ultrasound biomicroscopy (UBM) and the real number/size of SO droplets., Methods: An observational, cross-sectional study. Thirty-five RRD patients (19 males; 35 eyes) who underwent pars plana vitrectomy and SO injection were enrolled in the study. SO emulsification signs on UBM (endothelial deposits, floating droplets, ghost images, hyperoleon, anterior chamber angle impregnation, anterior iris surface impregnation, posterior iris surface impregnation and ciliary body impregnation) were acquired and quantified two days before SO removal, while real number/size of the SO droplets with a diameter in 0.4-20 μm in the first 2 mL washout fluid collected intraoperatively during SO removal were assessed by Coulter counter. Then the correlations between these values were explored., Results: The grade of tissue impregnation and hyperoleon area in the anterior chamber demonstrated significant correlations with the number of large (5-20 μm) SO droplets ( r  = 0.336; P  = 0.048 and r  = 0.350; P  = 0.039, respectively); however, no significant correlations were observed with the total number of droplets (0.4-20 μm)or the number of small (0.4-5 μm) droplets (both P  ＞ 0.05)., Conclusion: The UBM quantitative grade of tissue impregnation and hyperoleon area in the anterior chamber could provide a noninvasive overview of the actual degree of SO emulsification before SO removal surgery., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

In the development of ultrasound localization microscopy (ULM) methods, appropriate test beds are needed to facilitate algorithmic performance calibration. Here, we present the design of a new ULM-compatible microvascular phantom with a forked, V-shaped wall-less flow channel pair ( 250 μ m channel width) that is bifurcated at a separation rate of 50 μ m/mm. The lumen core was fabricated using additive manufacturing, and it was molded within a polyvinyl alcohol (PVA) tissue-mimicking slab using the lost-core casting method. Measured using optical microscopy, the lumen core's flow channel width was found to be 252 ± 15 μ m with a regression-derived flow channel separation gradient of 50.89 μ m/mm. The new phantom's applicability in ULM performance analysis was demonstrated by feeding microbubble (MB) contrast flow (1.67 to 167 μ L/s flow rates) through the phantom's inlet and generating ULM images with a previously reported method. Results showed that, with longer acquisition times (10 s or longer), ULM image quality was expectedly improved, and the variance of ULM-derived flow channel measurements was reduced. Also, at axial depths near the lumen's bifurcation point, the current ULM algorithm showed difficulty in properly discerning between the two flow channels because of the narrow channel-to-channel separation distance. Overall, the new phantom serves well as a calibration tool to test the performance of ULM methods in resolving small vasculature.

Objective: To investigate the relationship between corneal diameter and internal corneal span determined from angle-to-angle distance using ultrasound biomicroscopy (UBM) in an observational cross-sectional patient population comprised of 54 eyes (28 healthy control eyes, ages 0.1 to 11.3 years; 26 eyes with primary congenital glaucoma, ages 0.1 to 3.5 years) from 41 pediatric participants ages 0.1 to 11.3 years (mean age: 3±3 years, median age: 2 years)., Methods: Forty cornea photographs with reference ruler and 110 UBM images were obtained. Three observers measured horizontal and vertical corneal diameter and angle-to-angle distance in each cornea photo and UBM image using ImageJ and the average values were used. Main outcome measures were Pearson correlation coefficient, linear regression, mean difference between corneal diameter and angle-to-angle distance, and intra-class correlation coefficients among measurements from all three observers for each parameter., Results: Corneal diameter and angle-to-angle distance had a strong positive correlation horizontally (Pearson r = 0.89, p<0.001) and vertically (r = 0.93, p<0.001). Correlation was consistent regardless of presence of primary congenital glaucoma and participant age. Regression analysis demonstrated a linear relationship between the parameters for horizontal (CD = 0.99*AA+0.28, R2 = 0.81, p<0.001) and vertical (CD = 0.91 *AA+1.32, R2 = 0.85, p<0.001) dimensions. Overall, reliability was good-excellent, ranging from an ICC of 0.76 for vertical corneal diameter to 0.90 for horizontal angle-to-angle distance., Conclusions: Based on the strong positive correlation found between corneal diameter and angle-to-angle distance in our study population, UBM image analysis can be used to accurately estimate corneal diameter from angle-to-angle distance in children with healthy eyes and primary congenital glaucoma. UBM may provide a useful intraocular alternative for estimating corneal diameter and monitoring diseases that affect the cornea in infants and children, such as congenital glaucoma., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Kolosky et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Purpose: In the present study, we introduce human lacrimal gland imaging using an ultrasound biomicroscopy (UBM) with a soft cover and show their findings., Methods: The representative UBM findings of palpebral lobes in seven subjects (four with non-Sjögren dry eye syndrome, one with Sjögren syndrome, and two healthy subjects) were described in this study. To prolapse the palpebral lobe, the examiner pulled the temporal part of the upper eyelid in the superotemporal direction and directed the subject to look in the inferonasal direction. We scanned the palpebral lobes longitudinally and transversely using UBM. We used an Aviso UBM with a 50 MHz linear probe and ClearScan., Results: In UBM of two healthy subjects, the echogenicity of the lacrimal gland was lower than that of the sclera and homogeneous. But the parenchyma of a patient with Sjögren dry eye syndrome was quite inhomogeneous compared to the healthy subjects. In two patients with dry eye syndrome, we were able to observe some lobules in the parenchyma. We could find excretory ducts running parallel at the surface of the longitudinal section in some subjects. In the longitudinal UBM scan of a subject, we observed a tubular structure at a depth of 1,500 μm that was considered a blood vessel. It ran from the superonasal to the inferotemporal direction. In a subject, we observed a large cyst beneath the conjunctiva., Conclusions: Lacrimal gland imaging using UBM has both advantages of optical coherence tomography and sonography, and could be useful for evaluating dry eye syndrome.

Ultrasound localization microscopy (ULM) allows for the generation of super-resolved (SR) images of the vasculature by precisely localizing intravenously injected microbubbles. Although SR images may be useful for diagnosing and treating patients, their use in the clinical context is limited by the need for prolonged acquisition times and high frame rates. The primary goal of our study is to relax the requirement of high frame rates to obtain SR images. To this end, we propose a new time-efficient ULM (TEULM) pipeline built on a cutting-edge interpolation method. More specifically, we suggest employing Radial Basis Functions (RBFs) as interpolators to estimate the missing values in the 2-dimensional (2D) spatio-temporal structures. To evaluate this strategy, we first mimic the data acquisition at a reduced frame rate by applying a down-sampling (DS = 2, 4, 8, and 10) factor to high frame rate ULM data. Then, we up-sample the data to the original frame rate using the suggested interpolation to reconstruct the missing frames. Finally, using both the original high frame rate data and the interpolated one, we reconstruct SR images using the ULM framework steps. We evaluate the proposed TEULM using four in vivo datasets, a Rat brain (dataset A), a Rat kidney (dataset B), a Rat tumor (dataset C) and a Rat brain bolus (dataset D), interpolating at the in-phase and quadrature (IQ) level. Results demonstrate the effectiveness of TEULM in recovering vascular structures, even at a DS rate of 10 (corresponding to a frame rate of sub-100Hz). In conclusion, the proposed technique is successful in reconstructing accurate SR images while requiring frame rates of one order of magnitude lower than standard ULM.

Purpose: To investigate the correlation between swept-source anterior segment optical coherence tomography (AS-OCT) and ultrasound biomicroscopy (UBM) in congenital corneal opacity (CCO)., Methods: All children with unilateral or bilateral congenital corneal opacities who underwent examination under anesthesia (EUA) and anterior segment optical coherence tomography (AS-OCT) imaging from January 1, 2022, to December 31, 2022, were included. Main outcome measures were corneal and anterior segment evaluation and correlation of UBM and AS-OCT findings., Results: A total of 22 eyes of 15 patients were imaged using both technologies. The age at first EUA ranged from 11 days to 4 years. Different phenotypes were classified based on the clinical examination, UBM, and AS-OCT findings. Fourteen eyes were diagnosed with Peters anomaly, congenital corneal staphyloma was observed in 4 eyes, 2 eyes had coloboma, 1 eye had peripheral sclerocornea, and 1 eye was diagnosed with congenital primary aphakia. AS-OCT and UBM findings were closely correlated in 18 of 22 eyes (82%) but AS-OCT failed to provide detailed information in 4 eyes (18%) where UBM revealed more details., Conclusions: Although AS-OCT offers valuable preliminary data for initial assessment and counseling, it may not consistently provide precise assessments in all cases. Therefore, UBM should be considered for definitive evaluation., (Copyright © 2024 American Association for Pediatric Ophthalmology and Strabismus. Published by Elsevier Inc. All rights reserved.)

Purpose: To evaluate the anterior segment structures using ultrasound biomicroscopy (UBM) in primary congenital glaucoma (PCG) and explore their correlation with disease severity and surgical outcomes., Methods: Clinical information of PCG patients who underwent UBM prior to their first glaucoma surgeries from September 2014 to March 2021 were reviewed. The study included 214 UBM images of 154 PCG eyes and 60 fellow unaffected eyes. Anterior segment characteristics were analyzed. UBM parameters, including the iris thickness (IT) at variant distances from the pupil edge and iris root, anterior chamber depth (ACD), and pupil diameter (PD), were compared between two groups and their relationship with clinical factors and surgical outcomes were analyzed in PCG eyes., Results: PCG eyes had unclear scleral spur, thin iris, wide anterior chamber angle, deep anterior chamber, rarefied ciliary body, elongated ciliary processes, and abnormal anterior iris insertion. ITs were thinner, ACD was deeper, and PD was larger in PCG eyes than fellow unaffected eyes (all P < 0.001). In PCG eyes, thinner ITs correlated with bilateral involvement and earlier age at presentation, and larger PD correlated with earlier age at presentation (P = 0.030) and higher intraocular pressure (P < 0.001). Thinner IT2 (P = 0.046) and larger PD (P = 0.049) were identified as risk factors for surgical failure., Conclusion: UBM is a powerful technique to exam anterior segment structures in PCG. The anatomical features are associated with disease severity and surgical outcomes, providing essential clinical insights., (© 2023. The Author(s).)

Purpose: To evaluate the accuracy of the Reinstein formula with hand-held ultrasound biomicroscopy (UBM) measurements for sizing of the Implantable Collamer Lens (ICL)., Methods: A total of 107 myopic eyes of 57 patients implanted with the ICL were included in the study. The size of the ICL was selected based on the manufacturer's recommendations. Agreement between the vault predicted by the Reinstein formula and the vault measured postoperatively was analyzed with Bland-Altman plots., Results: A total of 95% and 81% of patients had a postoperative vault ranging from 150 to 1,000 and 250 to 750 μm, respectively. The mean vault predicted by the Reinstein formula and the postoperative vault in the current study were 580 ± 181 and 547 ± 200 μm, respectively. The size recommendations of the Reinstein formula and the formula provided by the manufacturer, the Kojima formula, and the Dougherty formula overlapped in 50%, 57%, and 49% of eyes, respectively., Conclusions: The results show that the Reinstein formula combined with a hand-held UBM provides reliable sizing predictions of the ICL. However, considering that robotic UBM measurements have demonstrated a narrower range of deviation in predicting vault depth in previous studies, a direct comparison study between robotic UBM and hand-held UBM measurements is necessary to fully assess the limitations of combining hand-held UBM with the Reinstein formula. [ J Refract Surg . 2024;40(3):e142-e147.] .

Ultrasound biomicroscopy (UBM) is an invaluable investigation for imaging anterior segment structures. Although it is operator-dependent and time consuming, unlike optical-based imaging techniques, it is able to image structures posterior to the iris, such as the zonules, ciliary body and part of the pars plana. It is especially useful in advanced cataracts, traumatic cataracts, subluxed lenses, posterior polar cataracts, and congenital and developmental anomalies affecting the anterior segment. It provides diagnostic information in eyes with complex cataracts or intraocular lens (IOL)-related pathology, and aids in surgical planning in order to minimise complications. In this review, we describe the UBM features of various lenticular pathologies and demonstrate its application in the diagnosis and surgical management of lens and IOL-related pathologies., (© 2024 Royal Australian and New Zealand College of Ophthalmologists.)

Significance: Photoacoustic Doppler flowmetry offers quantitative blood perfusion information in addition to photoacoustic vascular contrast for rectal cancer assessment., Aim: We aim to develop and validate a correlational Doppler flowmetry utilizing an acoustic resolution photoacoustic microscopy (AR-PAM) system for blood perfusion analysis., Approach: To extract blood perfusion information, we implemented AR-PAM Doppler flowmetry consisting of signal filtering and conditioning, A-line correlation, and angle compensation. We developed flow phantoms and contrast agent to systemically investigate the flowmetry's efficacy in a series of phantom studies. The developed correlational Doppler flowmetry was applied to images collected during in vivo AR-PAM for post-treatment rectal cancer evaluation., Results: The linearity and accuracy of the Doppler flow measurement system were validated in phantom studies. Imaging rectal cancer patients treated with chemoradiation demonstrated the feasibility of using correlational Doppler flowmetry to assess treatment response and distinguish residual cancer from cancer-free tumor bed tissue and normal rectal tissue., Conclusions: A new correlational Doppler flowmetry was developed and validated through systematic phantom evaluations. The results of its application to in vivo patients suggest it could be a useful addition to photoacoustic endoscopy for post-treatment rectal cancer assessment., (© 2024 The Authors.)

Purpose: Microglia, the brain's immune cells, play important roles in neuronal differentiation, survival, and death. The function of microglia is deeply related to the morphologies; however, it is too complex to observe conventionally and identify the condition of living microglia using optical microscopes. Herein, we proposed a new method to observe living cultured microglia and their reactivity to inflammation via the acoustic impedance mode of a scanning acoustic microscope., Methods: Primary cultured microglia collected from rat pups exposed to acetamiprid, an insecticide, in utero were observed with both acoustic interface impedance mode (C-mode) and transparent three-dimensional impedance mode (B-mode)., Results: We characterized microglia into four types based on the results obtained from acoustic impedance, cytoskeletal information, and laser confocal imaging. Biphasic acoustic observation using B-mode and C-mode gave us information regarding the dynamic morphologies of living microglia treated with adenosine triphosphate (ATP) (600 μmol/L), which reflects distress signals from inflamed neurons. Acetamiprid exposure induced microglia response even in the neonatal period. ATP stimulus altered the shape and thickness of microglia with a change in the bulk modulus of the cell. Three-dimensional alteration with ATP stimulus could be observed only after biphasic acoustic observation using B-mode and C-mode. This acoustic observation was consistent with confocal observation using anti-Iba-1 and P2Y12 immunocytochemistry., Conclusion: This study demonstrated the adequacy of using a scanning acoustic microscope in analyzing microglia's shape, motility, and response to inflammation., (© 2023. The Author(s).)

Purpose: This study evaluates the accuracy of modern intraocular lens (IOL) calculation formulas using axial length (AL) data obtained by ultrasound biometry (UBM) compared to the third-generation SRK/T calculator., Material and Methods: The study included 230 patients (267 eyes) with severe lens opacities that prevented optical biometry, who underwent phacoemulsification (PE) with IOL implantation. IOL power calculation according to the SRK/T formula was based on AL and anterior chamber depth obtained by UBM (Tomey Biometer Al-100) and keratometry on the Topcon KR 8800 autorefractometer. To adapt AL for new generation calculators - Barrett Universal II (BUII), Hill RBF ver. 3.0 (RBF), Kane and Ladas Super Formula (LSF) - the retinal thickness (0.20 mm) was added to the axial length determined by UBM, and then the optical power of the artificial lens was calculated. The mean error and its modulus value were used as criteria for the accuracy of IOL calculation., Results: A significant difference ( p =0.008) in the mean IOL calculation error was found between the formulas. Pairwise analysis revealed differences between SRK/T (-0.32±0.58 D) and other formulas - BUII (-0.16±0.52 D; p =0.014), RBF (-0.17±0.51 D; p =0.024), Kane (-0.17±0.52 D; p =0.029), but not with the LSF calculator (-0.19±0.53 D; p =0.071). No significant differences between the formulas were found in terms of mean error modulus ( p =0.238). New generation calculators showed a more frequent success in hitting target refraction (within ±1.00 D in more than 95% of cases) than the SRK/T formula (86%)., Conclusion: The proposed method of adding 0.20 mm to the AL determined by UBM allows using this parameter in modern IOL calculation formulas and improving the refractive results of PE, especially in eyes with non-standard anterior segment structure.

Objective: The goal of the work described here was to develop and assess a deep learning-based model that could automatically segment anterior chamber angle (ACA) tissues; classify iris curvature (I-Curv), iris root insertion (IRI), and angle closure (AC); automatically locate scleral spur; and measure ACA parameters in ultrasound biomicroscopy (UBM) images., Methods: A total of 11,006 UBM images were obtained from 1538 patients with primary angle-closure glaucoma who were admitted to the Eye Center of Renmin Hospital of Wuhan University (Wuhan, China) to develop an imaging database. The UNet++ network was used to segment ACA tissues automatically. In addition, two support vector machine (SVM) algorithms were developed to classify I-Curv and AC, and a logistic regression (LR) algorithm was developed to classify IRI. Meanwhile, an algorithm was developed to automatically locate the scleral spur and measure ACA parameters. An external data set of 1,658 images from Huangshi Aier Eye Hospital was used to evaluate the performance of the model under different conditions. An additional 439 images were collected to compare the performance of the model with experts., Results: The model achieved accuracies of 95.2%, 88.9% and 85.6% in classification of AC, I-Curv and IRI, respectively. Compared with ophthalmologists, the model achieved an accuracy of 0.765 in classifying AC, I-Curv and IRI, indicating that its high accuracy was as high as that of the ophthalmologists (p > 0.05). The average relative errors (AREs) of ACA parameters were smaller than 15% in the internal data sets. Intraclass correlation coefficients (ICCs) of all the angle-related parameters were greater than 0.911. ICC values of all iris thickness parameters were greater than 0.884. The accurate measurement of ACA parameters partly depended on accurate localization of the scleral spur (p < 0.001)., Conclusion: The model could effectively and accurately evaluate the ACA automatically based on fully automated analysis of UBM images, and it can potentially be a promising tool to assist ophthalmologists. The present study suggested that the deep learning model can be extensively applied to the evaluation of ACA and AC-related biometric risk factors, and it may broaden the application of UBM imaging in the clinical research of primary angle-closure glaucoma., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2023 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Amyloid consists of insoluble beta-fibrillar proteins with stable structures. The Congo red staining method for histologically detecting amyloid is unsuitable for quantitatively assessing amyloid fibers. Scanning acoustic microscopy (SAM) detects the attenuation of sound (AOS) through sections. This study aimed to clarify whether AOS values reflected the amount of amyloid fibril degradation in tissues. Formalin-fixed paraffin-embedded unstained sections of various types of amyloidosis were digested with different endopeptidases. The AOS images after digestion were observed over time via SAM. The corresponding Congo red-stained images were followed to identify the amyloid. The amyloid and nonamyloid portions were statistically examined over time to determine the changes in the AOS values. Most of the amyloid areas showed significantly different AOS values from nonamyloid portions before digestion and significantly decreased after digestion; these findings corresponded with the disappearance and waning of the Congo red staining in the light microscopic images. Some nonamyloid areas with high AOS masked the reduction in AOS in the amyloid areas. The method used in this study may help detect the amyloid quantity and determine the appropriate treatment method for removing amyloid deposits from tissues., (© 2023. The Author(s).)

Purpose: To investigate the ciliary body anatomy and position of the implantable collamer lens (ICL) in low-vault eyes and analyze factors related to insufficient vault., Setting: Zhongshan Ophthalmic Center, Guangzhou, China., Design: Retrospective case-control observational study., Methods: In this study, 73 eyes of 73 patients with an insufficient vault (<250 μm) were matched with 73 eyes with an ideal vault (250 to 750 μm). Ultrasound biomicroscopy was used to determine the ciliary body morphology and ICL position. The biometric parameters acquired by Scheimpflug tomography were compared. The correlation between the vault and these factors was analyzed, and the least absolute shrinkage and selection operator method was used to screen the risk factors for low vault., Results: The low-vault group had a steeper corneal curvature, thicker lens thickness (LT), higher crystalline lens rise, and shorter axial length (AL) (all P < .005). The ciliary process length (CPL) and maximum ciliary body thickness (CBTmax) were significantly smaller, and the trabecular-ciliary angle (TCA), iris-ciliary angle (ICA), and ciliary sulcus width (CSW) were significantly greater in the low-vault eyes (all P < .005). The low-vault group had more ICL haptics below the ciliary process, and TCA, ICA, CPL, CBTmax, CSW, and haptic position were related to the postoperative vault (all P < .05). CPL, AL, and LT were identified as predictors of a low vault., Conclusions: Malposition of ICL haptics behind the ciliary process is a risk factor for low vault. A shorter CPL, thicker LT, and shorter AL are significant risk factors for the postoperative low vault., (Copyright © 2023 Published by Wolters Kluwer on behalf of ASCRS and ESCRS.)

Background: Ultrasound biomicroscopy (UBM) is a noninvasive imaging modality that enables in-vivo visualization of the structures of the anterior segment of the eye. Unlike routine ophthalmic diagnostic ultrasound which uses frequencies of 5-10 MHz, UBM utilizes ultrasound frequencies in the range of 50-100 MHz. The high-frequency probes in UBM allows for higher resolution and better visualization of subsurface ocular structures, even in the presence of anatomic or pathological obscuration. UBM has qualitative as well as quantitative applications in various disorders affecting the anterior segment of the eye. Despite its huge importance, many clinicians lack in knowledge about the technique and its clinical usefulness. The current educational video aims to address this gap in knowledge by highlighting the technique and various clinical indications of UBM., Purpose: The purpose of this video is to demonstrate the technique of UBM and showcase its quantitative and qualitative implications and importance through various clinical cases., Synopsis: UBM is an imaging technique that assesses the depth of tissue structures by measuring the time delay of the returning ultrasound signal. This modality is capable of measuring the size of various structures within the eye, such as the cornea, iris, ciliary body, sclera, and the depth of the anterior and posterior chamber. To perform a UBM, a transducer is inserted into a specially designed eye cup filled with distilled water, creating a water bath environment. Axial and longitudinal scans can be performed in a similar fashion as in routine diagnostic B-scan ultrasound. Quantitative indications for UBM depicted in this video include measurements of corneal thickness, depth of the anterior chamber, and the width of the angle. The video also showcases how UBM can aid in the diagnosis and management of various anterior segment disorders like angle-closure glaucoma, plateau iris configuration, secondary glaucoma, and anterior uveitis with complicated cataract. Qualitative indications for UBM highlighted in this video include its role in intermediate uveitis, ocular hypotony, ocular surface tumors, cystic lesions of iris, and identifying the location and type of intraocular foreign bodies in the anterior segment based on the type of artifact seen. Additionally, the video shows the applications of UBM in scleral and episcleral pathologies., Highlights: This video will educate clinicians about the technique of UBM and showcase a bouquet of UBM findings in various case scenarios, helping one to better understand the potential of this modality in clinical practice., Video Link: https://youtu.be/F626TMbJXoU., Competing Interests: None

Background: There has not been a thoroughly reported study of the comparison between spectral domain-optical coherence tomography (SD-OCT) with both ultrasound biomicroscopy (UBM) and gonioscopy on the evaluation of the iridocorneal angle (ICA) in dogs., Objectives: To investigate the diagnostic value of SD-OCT for the early detection of narrowing ICA by comparing and assessing inter-device agreement in anterior chamber angle (ACA) measurements obtained by SD-OCT and UBM, and ICA evaluations by gonioscopy., Methods: A total of 28 eyes from 28 client-owned dogs with normal intraocular pressure were included for examination. The ACA and angle opening distance (AOD) were measured from the SD-OCT and UBM images, and gonioscopy images were analyzed using the ICA grade and ZibWest angle index., Results: The mean ACA and AOD for SD-OCT were 28.31° ± 5.37° and 658.42 ± 219.90 µm, and for UBM, 28.34° ± 5.82° and 859.29 ± 221.80 µm, respectively. The mean difference in ACA between the average values of SD-OCT and UBM measurements was 0.03° with a 95% limit of agreement (LoA) span of 16.2°, indicating positive agreement; that in AOD was 200.85 µm with a 95% LoA span of 1,110.95 µm, indicating poor agreement. The Pearson correlation coefficient of the ACA of SD-OCT and ZibWest indices of gonioscopy was 0.624, indicating strong agreement; that of UBM and gonioscopy was 0.43, indicating moderate agreement., Conclusions: SD-OCT is well tolerated by canine patients due to its non-contact method and might be an alternative option for early screening of ICA narrowing in clinical settings., Competing Interests: The authors declare no conflicts of interest., (© 2023 The Korean Society of Veterinary Science.)

Background: With ultrasound biomicroscopy (UBM), radial scans are taken by using a typical ciliary process, to show the details of the iridocorneal angle, the anterior surface of the ciliary body, and its relation to the posterior iris. Appositional closure represents potentially reversible contact between the peripheral iris and trabecular meshwork. The appositional closure can further be classified according to the configuration of iridotrabecular contact (ITC). UBM can be performed in dark and light conditions, which has been shown to be useful for detecting changes in iridocorneal angle configuration associated with dark and light., Purpose: To image ITC configuration in appositional angle closure and also image iridocorneal angle in dark and bright light room illumination. Synopsis: UBM demonstrates two types of ITC configuration in appositional closure which are, B-type and S-type. It can also demonstrate the presence of sinus of Mapstone in S-type of ITC., Highlights: UBM allows imaging of dynamic changes in the iris and shows that the degree of appositional angle closure is a dynamic process that can change rapidly depending on the lighting conditions., Video Link: https://youtu.be/tgN4SLyx6wQ., Competing Interests: None

Background: Ultrasound biomicroscopy (UBM) is a high-resolution ultrasound technique, which allows noninvasive, in vivo imaging of the ocular anterior segment structures., Purpose: This video gives a description of the identification of the iridocorneal angle structures in the cross-sectional view in a radial scan through a typical ciliary process and a guide toward measuring the angle parameters., Synopsis: UBM provides two-dimensional, grayscale images of the iridocorneal angle. The real-time image is displayed on a video monitor and can be recorded for qualitative and quantitative analysis. The angle parameters can be measured by in-built calipers in the machine software and manipulated by the examiner. Highlights: This video demonstrates UBM caliper positions as displayed on the monitor and marked by the examiner for the measurement of various anterior segment parameters of the eye., Video Link: https://youtu.be/WTXMupYoyww., Competing Interests: None

Background: Ultrasound biomicroscopy (UBM) is a high-resolution ultrasound technique that allows noninvasive, in vivo imaging of the ocular anterior segment structures. Before interpreting the UBM images of the diseased eyes, it is essential to understand the structures seen in the UBM image of the normal eye., Purpose: This video is a compilation of short video clips that gives description of identification of the anterior segment structures in the axial scan, a cross-sectional view through the anterior chamber angle region of a normal subject in a radial scan, and identification of ciliary processes in the transverse scan., Synopsis: UBM provides two-dimensional, grayscale images of the various anterior segment structures and allows all these structures to be imaged simultaneously, in their normal state, as they occur in the living eye. The real-time image is displayed on a video monitor and can be recorded for qualitative and quantitative analysis., Highlights: The video gives an overview of identification of normal anterior segment structures on UBM. Video link https://youtu.be/3KooOp2Cn30., Competing Interests: None

A female patient in her 20s presented with a bulbar conjunctival mass lesion that was diagnosed as nodular scleritis. It was treated with topical and oral steroids in another hospital. Imaging was done using anterior segment optical coherence tomography and ultrasound biomicroscopy, which helped to rule out scleritis and subconjunctival cysticercosis. Histopathology of the excision biopsy specimen revealed fungal filaments. Topical antifungals were started, and the condition resolved without recurrence after therapy. Although rare, infections should be considered in the differential diagnosis of conjunctival mass lesions. Imaging can help to rule out other entities and guide towards appropriate management., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

Purpose: The aim of the study was to compare and explore the agreement between the nomogram tool and ultrasound biomicroscopy (UBM) images method to calculate the ultrasound cycloplasty (UCP) probe model in Chinese glaucoma patients., Methods: Retrospective analysis of Chinese glaucoma patients who visited Zhongshan Ophthalmic Center in Guangzhou from January to December 2019 and were eligible for UCP surgery. Visual acuity, intraocular pressure (IOP), ocular axial length (AL), and horizontal corneal diameter (white to white [WTW]) were measured. UBM images with clear ciliary body imaging and AL and WTW data were sent to trained personnel for probe model measurements. The data calculated by both methods were analyzed using unweighted and weighted κ statistics. The level of agreement refers to Landis and Koch's guideline for the strength of agreement indicated with weighted κ values., Results: 1,061 eyes of 642 patients were involved, with a mean age of 61.66 ± 11.66 years. Their best-corrected visual acuity converted to logarithm of minimal-angle-of-resolution (logMAR) scores of -0.18-3.00 with a mean value of 0.69 ± 0.77. IOP was 22.0-60.0 mm Hg with a mean of 27.97 ± 5.66 mm Hg. The mean AL and WTW were 22.88 ± 1.33 (19.15-32.14) mm and 11.52 ± 0.49 (10.00-12.90) mm, respectively. The agreement between the two methods was fair (weighted κ = 0.299), matching in 62.86% of eyes (weighted κ = 0.299, κ = 0.264). The agreement in primary open angle glaucoma, acute primary angle-closure glaucoma, chronic primary angle-closure glaucoma, and secondary glaucoma patients was 60.85% (weighted κ = 0.336, κ = 0.301), 65.06% (weighted κ = 0.146, κ = 0.127), 62.26% (weighted κ = 0.204, κ = 0.184), and 57.97% (weighted κ = 0.332, κ = 0.280) of eyes, respectively., Conclusion: The agreement between UBM images and the nomogram tool to calculate the UCP probe model of Chinese patients is at a fair level. The nomogram tool prefers to use larger probes. Improvements to the nomogram tool, such as including data from more ethnic groups and being able to calculate separately for different types of glaucoma, are needed to improve accuracy. The inclusion of parameters or images from more directions of the eye may help measure probe models more accurately for both the nomogram tool and the UBM image measurement., (© 2023 The Author(s). Published by S. Karger AG, Basel.)

Quantitative acoustic microscopy (QAM) reconstructs two-dimensional (2D) maps of the acoustic properties of thin tissue sections. Using ultrahigh frequency transducers (≥ 100 MHz), unstained, micron-thick tissue sections affixed to glass are raster scanned to collect radiofrequency (RF) echo data and generate parametric maps with resolution approximately equal to the ultrasound wavelength. 2D maps of speed of sound, mass density, acoustic impedance, bulk modulus, and acoustic attenuation provide unique and quantitative information that is complementary to typical optical microscopy modalities. Consequently, many biomedical researchers have great interest in utilizing QAM instruments to investigate the acoustic and biomechanical properties of tissues at the micron scale. Unfortunately, current state-of-the-art QAM technology is costly, requires operation by a trained user, and is accompanied by substantial experimental challenges, many of which become more onerous as the transducer frequency is increased. In this chapter, typical QAM technology and standard image formation methods are reviewed. Then, novel experimental and signal processing approaches are presented with the specific goal of reducing QAM instrument costs and improving ease of use. These methods rely on modern techniques based on compressed sensing and sparsity-based deconvolution methods. Together, these approaches could serve as the basis of the next generation of QAM instruments that are affordable and provide high-resolution QAM images with turnkey solutions requiring nearly no training to operate., (© 2023. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

Acoustic-resolution photoacoustic micro- scopy (AR-PAM) system can provide 3-D images of facial tissues. The lateral resolution of AR-PAM depends on the numerical aperture (NA) of the acoustic lens and the central frequency of the ultrasonic transducer. There is a trade-off between resolution enhancement and imaging depth. The acoustic beam is tight in the acoustic focal plane but expands in the out-of-focus regions, deteriorating the resolution. High-NA AR-PAM has depth-variant resolution. Synthetic aperture focusing technique (SAFT) based on a virtual detector (VD) concept can compensate for the beam shape and improve the lateral resolution via beamforming. Although, beamforming can enhance the resolution but the lateral resolution in the focal plane is still limited by acoustic diffraction. Structured-illumination can shift the spatial spectrum of an image to low frequencies hence high-frequency contents can be reserved to overcome the diffraction limit. Conventional structured-illumination via using a three-phase-shifting method can improve the resolution by two folds. Here, a modified phase-shifting method is used to generate the second harmonic of the fringes and double the spectral shift. In this idea, higher frequency information compared to the three-phase shifting method can fall into the band-limited system response. The modified phase-shifting method expands the spatial bandwidth and increases the lateral resolution by five folds. The mathematical relations and the theory are discussed in the context. Tungsten filament result shows resolution improvement from 44.6 [Formula: see text] to 11.3 [Formula: see text] by the modified structured illumination. In vivo and ex vivo experimental results validate the system performance.

Background: The iridocorneal angle (ICA) is the major pathway of aqueous humour outflow from the anterior chamber of the eye. Ultrasound biomicroscopy (UBM) has been utilised to characterise the morphology of this drainage pathway in numerous species. UBM may allow for early recognition of aqueous humour outflow obstructions in horses, allowing for earlier recognition of risk for glaucoma, a vision-threatening and painful disease. UBM morphology of the normal equine ICA has yet to be described., Objectives: To determine the ultrasonographic morphology of the equine ICA by UBM in standing sedated horses., Study Design: In vivo experimental study., Methods: Thirty healthy adult horses underwent UBM of the ICA at four locations (superior, temporal, inferior, nasal) of each eye utilising standing sedation, topical anaesthesia and auriculopalpebral perineural anaesthesia. Anatomic structures were defined on ultrasound images through comparison to published histologic photomicrographs of the equine ICA., Results: Ultrasound imaging of the ICA at all four locations was easily performed in standing, sedated horses. High-resolution images of the ICA allowed for identification of the pectinate ligament, corneoscleral trabecular meshwork (TM), uveal TM and supraciliary TM., Main Limitations: Pupil size was midrange in all eyes, but was not strictly controlled. Lighting conditions not controlled. Various breeds included., Conclusion: In vivo UBM of the equine ICA is feasible and provides high-resolution images of the structures of the aqueous humour outflow pathway., (© 2022 EVJ Ltd.)

Background: To evaluate a new method of implantable collamer lens (ICL) sizing based on ultrasound biomicroscopy (UBM) video clips., Methods: This observational study included consecutive patients with myopia and myopic astigmatism scheduled for V4c toric ICL (TICL) implantation (STAAR) at Hangzhou MSK Eye Hospital (October 2020 to November 2020). Sulcus-to-sulcus (STS) distance, lens thickness (LT), and clinical refraction were measured preoperatively. The ZZ ICL formula (provides the predicted vault height and refraction based on TICL size, intraocular meridian, power, and eye parameters, including STS distance and LT) was used to select TICL size and predict vault height and residual refraction, which was also compared with the STAAR software recommended. Vault and residual refraction were measured at 3 months postoperatively., Results: The analysis included 168 eyes in 84 patients. Postoperative vault size was comparable to that predicted by the ZZ ICL formula (528 ± 193 vs. 545 ± 156 μm, P = 0.227). Vault prediction error (PE) by the ZZ ICL formula was within 100, 300, and 500 μm in 40.48%, 88.10%, and 100% of eyes, respectively. Spherical equivalent (SE) and absolute cylindrical refractive error were 0.36 ± 0.48 and 0.40 ± 0.31 D at 3 months postoperatively. The SE PE, absolute cylindrical PE, and percentages of eyes with an absolute cylindrical PE within ± 0.50 D and ± 1.00 D were lower for the ZZ ICL formula than for the STAAR software (P < 0.01)., Conclusions: Combining measurements obtained in UBM video clips with the ZZ ICL formula provides an effective method of sizing TICLs and predicting vault height and residual refractive error., (© 2022. The Author(s).)

Purpose: Primary congenital glaucoma (PCG) is a form of childhood glaucoma caused by maldevelopment of the anterior chamber. Disease severity differs greatly amongst patients. Ultrasound biomicroscopy (UBM) is a non-invasive technique that can visualize the anterior segment in infants in vivo. The purpose of this narrative review is to make an overview of the UBM data in PCG and study the applicability of UBM in characterizing the disease., Methods: An online search was performed on PubMed in December 2020. After a critical appraisal of the included articles, study and patient characteristics were summarized. The UBM measurements of the anterior segment in PCG of the different studies were analysed., Results: Six studies were included in this review. All were cross-sectional prospective studies. A total of 221 PCG eyes were examined. PCG eyes showed a larger trabecular iris angle, decreased iris thickness, narrower or absent Schlemm's canal and an increased zonular length compared to controls. Abnormal tissue membrane covering the trabecular meshwork and abnormal insertion of the iris and ciliary process were frequently found. The success rate of glaucoma surgery depended on the severity of anterior segment malformations found with UBM., Conclusion: Malformations of the anterior segment in PCG can be demonstrated by UBM in vivo. This imaging can help to characterize disease severity and might support surgical treatment decisions., (© 2021 The Authors. Acta Ophthalmologica published by John Wiley & Sons Ltd on behalf of Acta Ophthalmologica Scandinavica Foundation.)

Background/objective: Anterior segment optical coherence tomography (AS-OCT) and ultrasound biomicroscopy (UBM) are two non-invasive imaging techniques used for the measurement of tumour thickness in corneal and bulbar conjunctival tumours. Histopathology (HP), however, remains the gold standard for the measurement of tumour thickness. The aim of this study was to determine whether AS-OCT and UBM are as accurate as HP for measuring tumour thickness., Methods: Forty-two corneal and bulbar conjunctival tumours were imaged using AS-OCT and UBM. Images were assessed and tumour thickness was measured. Eleven patients subsequently underwent surgical excision. All specimens were measured during histopathological analysis. The correlation of the thickness measurement on HP to AS-OCT and UBM was then statistically analysed. In cases where the tumour was not excised, thickness measurement comparisons between AS-OCT and UBM were analysed., Results: AS-OCT and UBM measurements of tumour thickness were found to be significantly positively correlated (p=<0.001), as were UBM and HP thickness measurements (p=0.031). HP and AS-OCT measurements, however, only showed a mild but non-significant positive correlation., Conclusion: Both AS-OCT and UBM are useful techniques to image and measure the thickness of corneal and conjunctival bulbar tumours. While AS-OCT provides better details than UBM, it was more limited in visualising the posterior boundary of the tumour, particularly in malignant tumours. While thickness measurements of both methodologies were correlated, neither should yet be considered as replacements to the gold standard of HP., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

Clinical Relevance: Ultrasound biomicroscopy is an objective method for assessing changes in anterior segment biometry. There is a paucity of data on the reliability of this method. A reliable method for assessing anterior segment changes during physiologically driven accommodation can be a useful tool for clinicians, researchers, and industry., Background: To assess the test-retest reliability of ultrasound biomicroscopy for measurements of change in anterior chamber depth during a distance to near fixation task in pseudophakic subjects., Methods: Subjects were adults with monofocal intraocular lenses implanted in both eyes who completed a 6-month post-operative period and had monocular uncorrected distance visual acuity of 6/15 (0.4 logMAR) or better. The change in anterior chamber depth during a distance to near fixation task was measured with a 35-MHz VuMAX HD ultrasound biomicroscopy device (Sonomed Escalon, New Hyde Park, NY) during two separate visits. An asymmetrical vergence paradigm allowed evaluation of anterior segment biometry at 22-µm axial resolution in one eye, while the fellow eye fixated on the target. To assess the test-retest reliability, 2-sided 95% CI from a paired t test was calculated for the difference in anterior chamber depth change from distance to near between visits., Results: The mean (standard deviation) near-focused anterior chamber depth measured by ultrasound biomicroscopy was 4.331 (0.237) and 4.333 (0.241) mm at visits 1 and 2, respectively. In response to a change in fixation from distance (4 m) to near (40 cm), the mean anterior chamber depth change was -0.012 (0.038) and 0.003 (0.039) mm at visits 1 and 2, respectively. Analysis of the difference in the change in anterior chamber depth between visits was -0.015 mm (95% CI, -0.035 to 0.003)., Conclusion: Ultrasound biomicroscopy is a repeatable, objective method for assessing change in anterior segment biometry during physiological changes in fixation from distance to near.

Objective: To compare the measurements of iridocorneal angle parameters between spectral domain optical coherence tomography (SD-OCT) and ultrasound biomicroscopy (UBM) in dogs., Animals Studied and Procedures: A total of 47 eyes of dogs were scanned at the temporal limbus using SD-OCT and UBM. Iridocorneal angle (ICA) and angle opening distance (AOD) were measured from the obtained images accordingly. The intra-observer and inter-observer reproducibility were evaluated using the intraclass correlation coefficient. To evaluate intra-observer reproducibility, measurements of the first and second grading from the first examiner were compared. To evaluate inter-observer reproducibility, measurements between the two examiners were compared. Agreement between ICA and AOD for SD-OCT and UBM was evaluated using Bland-Altman plots., Results: In the first grading, the mean ICA and AOD for SD-OCT were 31.4 ± 6.4° and 641.4 ± 270.8 µm, respectively. The mean ICA and AOD for UBM were 32.0 ± 4.8° and 700.4 ± 238.8 µm, respectively. For ICA and AOD measurements, intra-observer reproducibility was excellent for both devices, whereas inter-observer reproducibility was excellent for SD-OCT and good for UBM. The mean difference in ICA between SD-OCT and UBM was 0.6° with a limit of agreement (LoA) span of 18.9°. The mean difference in AOD between SD-OCT and UBM was 58.9 µm with a LoA span of 804.4 µm., Conclusions: Spectral domain optical coherence tomography is an effective non-contact imaging modality for the evaluation of canine iridocorneal angle parameters in a clinical setting. Reproducibility of measurements obtained is comparable or superior to UBM, but values obtained by SD-OCT and UBM for AOD are not interchangeable between devices., (© 2021 American College of Veterinary Ophthalmologists.)

Objective: To determine corneal thickness (CT) and axial anterior chamber depth (ACD) using ultrasound biomicroscopy (UBM) in normal adult horses. To compare corneal thickness measurements between UBM and ultrasonic pachymetry., Animals Studied: Sixty eyes of 30 healthy adult horses aged 8-24 years., Procedures: Ultrasonic pachymetry (velocity of 1640 m/s) was utilized to obtain measurements of the central, superior, temporal, inferior, and nasal cornea. Triplicate images of the same corneal locations were acquired using UBM (50 MHz). Images of the axial anterior chamber were used to measure ACD. Intraocular pressure (IOP) was estimated using rebound tonometry, and axial globe length was measured using ultrasonographic biometry., Results: CT (mean ± SD µm) measured by UBM was 854 ± 61 (central), 994 ± 58 (superior), 930 ± 57 (temporal), 979 ± 55 (inferior), and 898 ± 48 (nasal). CT measured by UBM was greater than that measured by ultrasonic pachymetry at all locations and was statistically significant at all locations except inferior (p = 0.0006-0.048). No sex nor age effect was detected for CT at any location. The repeatability of ultrasonic pachymetry was superior to that of UBM. Mean ± SD ACD was 5.74 ± 0.41 mm. A weak positive correlation was identified between central CT and IOP and between central CT and axial globe length., Conclusions: Normal data for CT and ACD of the adult horse obtained using UBM are provided. CT determined by UBM was greater relative to pachymetry at all corneal locations., (© 2022 American College of Veterinary Ophthalmologists.)

Purpose: To compare the accuracy of anterior segment optical coherence tomography (AS-OCT) and ultrasound biomicroscopy (UBM) in localizing rectus muscle insertions., Methods: The study was performed on 27 patients (39 rectus muscles) who required primary or secondary surgery. Using caliper function in the AS-OCT and UBM software, the distance from the insertion site to the anterior chamber angle was measured. The actual muscle insertion distance from limbus was considered as the measured distance plus 1 mm. The measurements by UBM and AS-OCT were compared with intraoperative measurements and with each other., Results: AS-OCT and UBM were performed on 13 medial rectus, 24 lateral rectus, and 2 superior rectus muscles. Ninety two percent of UBM measurements (36 muscles) were within 1 mm, one was within 1-1.5 mm, and 2 were within 1.5-2 mm of surgery measurements. Eighty five percent of AS-OCT measurements (33 muscles) were within 1 mm, 5 were within 1-1.5 mm, and one was within 1.5-2 mm of surgery measurements. In all cases, the mean absolute error of the UBM (0.54 ± 0.44) and AS-OCT (0.51 ± 0.36) showed no significant difference (p = .76)., Conclusion: AS-OCT and UBM can be used interchangeably to localize rectus muscle insertions and showed good agreement with intraoperative measurements.

Objective: The objective is to compare parameters related to lens position measured using anterior segment optical coherence tomography (AS-OCT) and ultrasound biomicroscopy (UBM) in patients with senile cataract and perform a consistency analysis., Methods: This prospective study included 102 patients (102 eyes) scheduled for simple cataract surgery. Among the total patients, 44 were men, and 58 were women. AS-OCT (sitting) and UBM (lying) were used to measure the anterior chamber depth (ACD) in horizontal and vertical orientations and the iris-lens contact distance (ILCD) and iris-lens angle (ILA) in inferior, superior, nasal, and temporal quadrants. Paired-sample t -test was used to compare ACD, ILCD, and ILA measurements of the two methods, while Pearson's linear correlation and Bland-Altman analyses were used to analyze the correlation and consistency of the two results., Results: The horizontal (2.499 ± 0.464 mm) and vertical (2.531 ± 0.463 mm) ACD measured using AS-OCT and the horizontal (2.556 ± 0.467 mm) and vertical (2.563 ± 0.479 mm) ACD measured using UBM were significantly different ( P < 0.001); moreover, the results showed good correlation and agreement. A significant difference was observed between the two methods in terms of ILCD measured in inferior, superior, nasal, and temporal quadrants ( P < 0.001), and a significant correlation was found between measurements of both methods ( P < 0.001). Approximately 3.92% (4/102), 0.98% (1/102), 3.92% (4/102), and 2.94% (3/102) of points were outside the 95% limits of agreement in the four quadrants, respectively, and the agreement of the results was good. ILA measured using both methods differed in inferior, superior, nasal, and temporal quadrants ( P =0.003, 0.011, 0.001, 0.001, respectively), and the correlation was good ( P < 0.001). The percentage of points outside the 95% limit was higher in inferior, superior, nasal, and temporal quadrants (4.90% (5/102), 5.88% (6/102), 5.88% (6/102), and 6.86% (7/102)) with poor agreement of the results., Conclusions: The correlation between AS-OCT and UBM in terms of measuring lens position-related parameters was good, but the agreement was unstable. The differences in measurement position (sitting and supine) and/or measurement methods (optics and ultrasound) may lead to variability in results., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2022 Zhiying Yu et al.)

Lung tissue stiffness is altered with aging. Quantitatively evaluating lung function is difficult using a light microscope (LM) alone. Scanning acoustic microscope (SAM) calculates the speed-of-sound (SOS) using sections to obtain histological images by plotting SOS values on the screen. As SOS is positively correlated with stiffness, SAM has a superior characteristic of simultaneously evaluating tissue stiffness and structure. SOS images of healthy bronchioles, arterioles, and alveoli were compared among young, middle-aged, and old lung sections. Formalin-fixed, paraffin-embedded (FFPE) sections consistently exhibited relatively higher SOS values than fresh-frozen sections, indicating that FFPE became stiffer but retained the relative stiffness reflecting fresh samples. All lung components exhibited gradually declining SOS values with aging and were associated with structural alterations such as loss of smooth muscles, collagen, and elastic fibers. Moreover, reaction to collagenase digestion resulted in decreased SOS values. SOS values of all components were significantly reduced in young and middle-aged groups, whereas no significant reduction was observed in the old group. Protease damage in the absence of regeneration or loss of elastic components was present in old lungs, which exbited dilated bronchioles and alveoli. Aging lungs gradually lose stiffness with decreasing structural components without exposure to specific insults such as inflammation., Competing Interests: The authors have declared that no competing interests exist.

Background: Advances in MRI coil technology and increased availability of high-field MRI in veterinary medicine enable the acquisition of images of increasingly high spatial resolution while preserving signal-to-noise ratio.The purpose of the present study was to compare 3T high-resolution magnetic resonance imaging (HR-MRI) with ultrasound (US) and ultrasound biomicroscopy (UBM) in the normal canine eye, to assess its potential to depict normal ocular anatomy., Results: HR-MRI was compared with US and UBM in 10 eyes from 10 healthy beagle dogs. Ocular structures (cornea, anterior chamber, iridocorneal angle, iris, lens, ciliary body, choroid, vitreous body, posterior wall of the eye, optic nerve and optic nerve sheath, extraocular muscles) were assessed subjectively and central corneal thickness (CCT), anterior chamber depth (ACD), aqueous depth (AQD), anteroposterior, mediolateral and dorsoventral lens diameter (APLD, MLLD, DVLD), anteroposterior diameter of the globe including and excluding the scleroretinal rim (APDSRR, APD), vitreous chamber depth (VCD) and optic nerve sheath diameter (ONSD) were measured in HR-MRI and in US. Optic nerve diameter (OND) was measured in HR-MRI. HR-MRI and UBM appearance of the anterior segment were subjectively compared. Detailed reference high-resolution MRI images of normal eyes of Beagle dogs are provided., Conclusions: HR-MRI allowed assessment of all structures identified with US and UBM. The MRI examinations were performed under general anesthesia with the addition of a neuromuscular blocking agent, while US and UBM examinations were performed in conscious animals. Visibility of the entire ocular wall, the lens, the structures caudal to the ciliary body and the optic nerve and its sheath was superior with HR-MRI. HR-MRI allowed the distinction of retina, choroid and sclera, and the delineation of structures not previously identified in canine eyes with MRI, including Tenon's capsule and the sub-Tenon's space.Plane selection was more accurate with HR-MRI compared to US. In general, the range of measurements was narrower for MRI than for US. CCT, AQD, APLD, MLLD, APD, APDSRR and ONSD differed significantly between HR-MRI and US, respectively (p = 0.005-0.027).Micro-MRI may be useful for the assessment of ocular pathologies in the future., (© 2022. The Author(s).)

Herein, we propose a method to estimate the reflection coefficient of the ultrasonic wave transmitted onto an object and to display this with acoustic impedance distribution. The observation targets were glial cells, which have a rigid cytoskeleton and spread out well on a culture substrate. A reflection coefficient derived only from the cells was then obtained using a deconvolution process. In the conventional method, the deconvolution process that was performed only in the frequency domain would cause an error in the reconstructed signal, and it formed an artifact when the result was converted into the acoustic impedance image. To solve this problem, two types of deconvolution techniques were applied in either the full frequency or time-frequency domain. The results of both methods were then compared. Since the characteristic acoustic impedance is a physical property substantially equivalent to the bulk modulus, it can be considered that the internal elastic parameter is thus estimated. An analysis of the nucleus based on its position in the acoustic impedance image was then performed. The results indicated that the proposed time-frequency domain deconvolution method is able to maintain the structure of the cell, while the cell itself is free from unwanted artifacts. The nucleus was also estimated to be located toward the center of the cell, with lower acoustic impedance value than the cytoskeleton. The results of this study could contribute to establishing a method for monitoring the internal condition of cultured cells in regenerative medicine and drug discovery., (Copyright © 2021 Elsevier B.V. All rights reserved.)