Non-telecentric photoacoustic microscopy for multi-scale imaging.

Optical-resolution photoacoustic microscopy (OR-PAM) excels in precisely imaging a biological tissue based on absorption contrast. However, existing OR-PAMs are confined by fixed compromises between spatial resolution and field of view (FOV), preventing the integration of large FOV and local high-resolution within one system. Here, we present a non-telecentric OR-PAM (nTC-PAM) that empowers efficient adaptation of FOV and spatial resolution to match the multi-scale requirement of diverse biological imaging. Our method allows for a large-scale transformation in FOV and even surpassing the nominal FOV of the objective with minimal marginal degradation of the lateral resolution. We demonstrate the advantage of nTC-PAM through multi-scale imaging of the leaf phantom, mouse ear, and cortex. The results reveal that nTC-PAM can switch the FOV and spatial resolution to meet the requirements of different biological tissues, such as large-scale imaging of the whole cerebral cortex and high-resolution imaging of microvascular structures in local brain regions.

Speckle Variance Photoacoustic Microscopy for Microhemodynamic Imaging.

Relying on the strong optical absorption of hemoglobin to pulsed laser energy, photoacoustic microscopy provides morphological and functional information on microvasculature label-freely. Here, we propose speckle variance photoacoustic microscopy (SV-PAM), which harnesses intrinsic imaging contrast from temporal-varied photoacoustic signals of moving red blood cells in blood vessels, for recovering three-dimension hemodynamic images down to capillary-level resolution within the microcirculatory tissue beds in vivo. Calculating the speckle variance of consecutive photoacoustic B-scan frames acquired at the same lateral position enables accurate identification of blood perfusion and occlusion, which provides interpretations of dynamic blood flow in the microvasculature, in addition to the microvascular anatomic structures. We demonstrate high-resolution hemodynamic imaging of vascular occlusion and reperfusion in the microvasculature of mice ears in vivo. The results suggest that our SV-PAM is potentially invaluable for biomedical hemodynamic investigations, for example, imaging ischemic stroke and hemorrhagic stroke.

Lista de verificación para la evaluación de imágenes de resonancia magnética en patología otológica / Checklist for the evaluation of magnetic resonance imaging in otological pathology

Los especialistas en Otorrinolaringología debemos estar familiarizados con los estudios radiológicos que nos permiten el diagnóstico de las distintas patologías otológicas. La resonancia magnética es un complemento a la tomografía computarizada, que permite una mejor evaluación de los tejidos blandos, lo cual contribuye al diagnóstico diferencial de las lesiones ocupantes de espacio localizadas en el hueso temporal y base de cráneo lateral. Es, además, la técnica de elección para la valoración del oído interno y de las estructuras anatómicas situadas en el ángulo pontocerebeloso. En este artículo presentamos una lista de verificación de resonancia magnética del oído con distintos apartados que permitirán hacer una revisión sistemática de todas las estructuras de interés en la práctica otológica, evaluables mediante esta técnica de imagen, así como las secuencias y cortes de elección en cada caso. (AU)

Otolaryngology specialists must be familiar with radiological studies that allow the diagnosis of different otological pathologies. Magnetic resonance imaging is a complement to computed tomography, which allows a better evaluation of soft tissues and contributes to the differential diagnosis of space-occupying lesions located in the temporal bone and lateral skull base. It is also the technique of choice for the evaluation of the inner ear and the anatomical structures located in the cerebellopontine angle. In this article we present a checklist for magnetic resonance imaging of the ear with different sections that will allow a systematic review of all structures of interest in otological practice, as well as the preferred sequences for each situation (AU).

Relevant anatomical details in rats regarding otologic research / Detalles anatómicos relevantes en ratas para la investigación otológica

SUMMARY: The objective of the study was to provide a detailed anatomical description of the rat's ear anatomy that will prove insightful to different experimental otologic surgical procedures regardless of scope. Three male Wistar rats were enrolled in the study. Candidates were screened for systemic and otologic pathology. External ear canal endoscopy was carried out with a 30˚ rigid endoscope through an image capture system. Middle ear anatomical elements were analyzed under stereomicroscopy. 3D computer tomography image reconstruction was realized with a micro-CT to describe the anatomy. Image data from all three rats were analyzed. Anatomical annotations and surgical exposure recommendations were added for key elements. The most relevant images from all three rats were selected for representation. Detailed visualization of the structural elements of the tympanic cavity were clearly visible: promontory, round window, stapedial artery, stapes, incus, and tympanic membrane were all constant findings. We describe a step wise ventral surgical approach of the middle and inner ear for which we found that the clavotrapezius muscle was a reliable landmark. For the transtympanic approach the endoscopic transcanal access was an easy and reliable method for which a detailed anatomical representation was depicted. Further, anatomical similarities to humans were observed by stereomicroscopy and Micro-CT imaging reiterating that the rat model is suitable for otologic research. The endoscopic approach to the tympanic membrane is comfortable and less expensive than a microscope. The tendon of the clavotrapezius muscle can be a reliable landmark for discovering the tympanic bulla when considering a ventral approach. 3D Micro-CT reconstruction allows intact evaluation of the samples, simultaneously being a diagnostic and also a learning tool.

RESUMEN: El objetivo de este trabajo fue proporcionar una descripción anatómica detallada de la anatomía del órgano vestíbulococlear de la rata que resultará útil para diferentes procedimientos quirúrgicos otológicos experimentales, independientemente del alcance. En el estudio se usaron tres ratas Wistar macho. Los ejemplares fueron evaluados por patología sistémica y otológica. La endoscopía del meato acústico externo se realizó con un endoscopio rígido de 30˚ a través de un sistema de captura de imágenes. Los elementos anatómicos del oído medio se analizaron bajo estereomicroscopía. La reconstrucción de la imagen de tomografía computarizada en 3D se realizó con un micro-CT para describir la anatomía. Se analizaron los datos de imagen de las tres ratas. Se agregaron anotaciones anatómicas y recomendaciones de exposición quirúrgica para elementos clave. Las imágenes más relevantes de las tres ratas fueron seleccionadas para su representación. La visualización detallada de los elementos estructurales de la cavidad timpánica era claramente visible: promontorio timpánico, ventana coclear, arteria estapedial, estapedio, yunque y membrana timpánica eran hallazgos constantes. Describimos un abordaje quirúrgico ventral escalonado del oído medio e interno para el cual encontramos que el músculo clavotrapecio era un punto de referencia confiable. Para el abordaje transtimpánico, el acceso transcanal endoscópico fue un método fácil y confiable para el cual se describió una representación anatómica detallada. Además, se observaron similitudes anatómicas con los humanos mediante estereomicroscopía e imágenes Micro-CT, lo que reitera que el modelo de rata es adecuado para la investigación otológica. El abordaje endoscópico de la membrana timpánica es cómodo y menos costoso que un microscopio. El tendón del músculo clavotrapecio puede ser un punto de referencia fiable para descubrir la bulla timpánica cuando se considera un abordaje ventral. La reconstrucción 3D Micro- CT permite la evaluación intacta de las muestras, siendo simultáneamente una herramienta de diagnóstico y también de aprendizaje.

Prenatal diagnosis of auriculocondylar syndrome with a novel missense variant of GNAI3: a case report.

BACKGROUND: Auriculocondylar syndrome (ACS) is a rare disorder characterized by micrognathia, mandibular condyle hypoplasia, and auricular abnormalities. Only 6 pathogenic variants of GNAI3 have been identified associated with ACS so far. Here, we report a case of prenatal genetic diagnosis of ACS carrying a novel GNAI3 variant. CASE PRESENTATION: A woman with 30 weeks of gestation was referred to genetic counseling for polyhydramnios and fetal craniofacial anomaly. Severe micrognathia and mandibular hypoplasia were identified on ultrasonography. The mandibular length was 2.4 cm, which was markedly smaller than the 95th percentile. The ears were low-set with no cleft or notching between the lobe and helix. The face was round with prominent cheeks. Whole-exome sequencing identified a novel de novo missense variant of c.140G > A in the GNAI3 gene. This mutation caused an amino acid substitution of p.Ser47Asn in the highly conserved G1 motif, which was predicted to impair the guanine nucleotide-binding function. All ACS cases with GNAI3 mutations were literature reviewed, revealing female-dominated severe cases and right-side-prone deformities. CONCLUSION: Severe micrognathia and mandibular hypoplasia accompanied by polyhydramnios are prenatal indicators of ACS. We expanded the mutation spectrum of GNAI3 and summarized clinical features to promote awareness of ACS.

Sonographic measurement of ear length among normal fetuses of pregnant Igbo women in port Harcourt, Nigeria.

BACKGROUND: Fetal ear length measurement has been associated with some clinical values: sonographic marker for chromosomal aneuploidy and for biometric estimation of fetal gestational age. OBJECTIVES: To establish a baseline reference value for fetal ear length and to assess relationship between fetal ear length and gestational age. METHODS: Ear length measurements were obtained prospectively from fetuses in 551 normal singleton pregnancies of 15 to 41 weeks gestation. Normal cases were defined as normal sonographic findings during examination plus normal infant post-delivery. The relationship between gestational age (GA) in weeks and fetal ear length (FEL) in millimeters were analyzed by simple linear regression. Correlation of FEL measurements with GA, biparietal diameter (BPD), Head circumference (HC), Abdominal Circumference (AC), Femur Length (FL) and maternal age (MA) were also obtained. RESULTS: Linear relationships were found between FEL and GA (FEL=0.872GA-2.972). There was a high correlation between FEL and GA (r = 0.837; P = .001). Good linear relationship and strong positive correlation were demonstrated between FEL and BPD, AC, HC, and FL (p<0.05). CONCLUSION: The result of this study provides normal baseline reference value for FEL. The study also showed good linear relationship and good correlation between FEL and fetal biometric measurements.

Comparative study of optical coherence tomography angiography algorithms for rodent retinal imaging.

Optical coherence tomography angiography (OCTA) is a functional extension of optical coherence tomography for non-invasive in vivo three-dimensional imaging of the microvasculature of biological tissues. Several algorithms have been developed to construct OCTA images from the measured optical coherence tomography signals. In this study, we compared the performance of three OCTA algorithms that are based on the variance of phase, amplitude, and the complex representations of the optical coherence tomography signals for rodent retinal imaging, namely the phase variance, improved speckle contrast, and optical microangiography. The performance of the different algorithms was evaluated by comparing the quality of the OCTA images regarding how well the vasculature network can be resolved. Quantities that are widely used in ophthalmic studies including blood vessel density, vessel diameter index, vessel perimeter index, vessel complexity index were also compared. Results showed that both the improved speckle contrast and optical microangiography algorithms are more robust than phase variance, and they can reveal similar vasculature features while there are statistical differences in the calculated quantities.

Photoacoustic-fluorescence microendoscopy in vivo.

A miniature endoscope capable of imaging multiple tissue contrasts in high resolution is highly attractive, because it can provide complementary and detailed tissue information of internal organs. Here we present a photoacoustic (PA)-fluorescence (FL) endoscope for optical-resolution PA microscopy (PAM) and FL microscopy (FLM). The endoscope with a diameter of 2.8 mm achieves high lateral resolutions of 5.5 and 6.3 µm for PAM and FLM modes, respectively. In vivo imaging of zebrafish larvae and a mouse ear is conducted, and high-quality images are obtained. Additionally, in vivo endoscopic imaging of a rat rectum is demonstrated, showing the endoscopic imaging capability of our endoscope. By providing dual contrasts with high resolution, the endoscope may open up new opportunities for clinical endoscopic imaging applications.

Inner ear salivary gland choristoma extending to the middle ear with congenital profound hearing loss and facial palsy: a case report.

BACKGROUND: Salivary gland choristoma (SGCh) is a rare benign tumor reported in several unusual sites, such as the gastrointestinal tract, the optic nerve, and the internal auditory canal, but never reported in the inner ear. CASE PRESENTATION: An 8-year-old girl with a history of left profound congenital hearing loss presented to us with ipsilateral progressive severe facial nerve palsy (House-Brackmann Grade VI). The left tympanic membrane was swollen with a pulsatile tumor. Radiological investigations revealed a multilocular tumor in the inner ear extending into the middle ear and internal auditory canal (IAC). We performed a partial resection of the tumor by transmastoid approach to preserve the anatomical structure of the facial nerve. The tumor was pathologically diagnosed as SGCh. Two years after surgery, her facial function recovered to House-Brackmann Grade II and the residual tumor did not show regrowth on MRI. CONCLUSIONS: Although the natural course of this rare tumor is unknown, a partial resection is an acceptable treatment procedure when functional recovery of the facial nerve is anticipated.

A novel missense variant of the GNAI3 gene and recognisable morphological characteristics of the mandibula in ARCND1.

Auriculocondylar syndrome (ARCND) is an autosomal monogenic disorder characterised by external ear abnormalities and micrognathia due to hypoplasia of the mandibular rami, condyle and coronoid process. Genetically, three subtypes of ARCND (ARCND1, ARCND2 and ARCND3) have been reported. To date, five pathogenic variants of GNAI3 have been reported in ARCND1 patients. Here, we report a novel variant of GNAI3 (NM_006496:c.807C>A:p.(Asn269Lys)) in a Japanese girl with micrognathia using trio-based whole exome sequencing analysis. The GNAI3 gene encodes a heterotrimeric guanine nucleotide-binding protein. The novel variant locates the guanine nucleotide-binding site, and the substitution was predicted to interfere with guanine nucleotide-binding by in silico structural analysis. Three-dimensional computer tomography scan, or cephalogram, displayed severely hypoplastic mandibular rami and fusion to the medial and lateral pterygoid plates, which have been recognised in other ARCND1 patients, but have not been described in ARCND2 and ARCND3, suggesting that these may be distinguishable features in ARCND1.

Deep learning differentiates between healthy and diabetic mouse ears from optical coherence tomography angiography images.

We trained a deep learning algorithm to use skin optical coherence tomography (OCT) angiograms to differentiate between healthy and type 2 diabetic mice. OCT angiograms were acquired with a custom-built OCT system based on an akinetic swept laser at 1322 nm with a lateral resolution of ∼13 µm and using split-spectrum amplitude decorrelation. Our data set consisted of 24 stitched angiograms of the full ear, with a size of approximately 8.2 × 8.2 mm, evenly distributed between healthy and diabetic mice. The deep learning classification algorithm uses the ResNet v2 convolutional neural network architecture and was trained on small patches extracted from the full ear angiograms. For individual patches, we obtained a cross-validated accuracy of 0.925 and an area under the receiver operating characteristic curve (ROC AUC) of 0.974. Averaging over multiple patches extracted from each ear resulted in the correct classification of all 24 ears.

The use of near infrared fluorescence imaging with indocyanine green for vascular visualization in caudal auricular flaps in two cats.

OBJECTIVES: To describe the use of near infrared fluorescence (NIRF) imaging with indocyanine green (ICG) for vascular visualization in two caudal auricular axial pattern flaps (APF). ANIMALS: Two client-owned cats with upper eyelid masses. STUDY DESIGN: Clinical case report. METHODS: Wide surgical excision with enucleation was performed by using a caudal auricular APF for closure. Flap margins and perforating artery location were approximated with anatomical landmarks. The caudal auricular artery origin was then visualized percutaneously by using an exoscope with NIRF camera and light source after a single 2.5-mg dose of IV ICG. Margins were adjusted as required. The flaps were routinely elevated with continued intraoperative visualization of the artery and rotated to complete closure. RESULTS: After IV ICG administration, fluorescence was initially visualized after 15 to 18 seconds and remained visible for up to 26 minutes. The achieved visualization led to flap margin adjustments in cat 1. Both cats recovered with minimal flap congestion, excellent hair regrowth, and no long-term complications (>186 days). Cat 1 experienced 100% flap survival. Cat 2 experienced 10% partial thickness flap necrosis, but revision was not required, and the flap was healed at recheck 85 days postoperatively. CONCLUSION: The use of ICG for APF vessel visualization prior to and during flap elevation resulted in transcutaneous visualization of the perforating vessel and improved awareness of vessel location intraoperatively. These two cats experienced excellent flap survival without major complications. This report highlights the potential benefits of ICG NIRF in APF for animals undergoing reconstructive surgery.

Intravital Imaging of Vascular Permeability by Two-Photon Microscopy.

The regulation of vascular permeability is critical in inflammation. It controls the distribution of water and plasma contents such as immunoglobulins in peripheral tissues. To regulate allergic diseases, it is important to study vascular biology especially in inflammation. Since the vascular permeability changes in minutes upon the exposure to proinflammatory mediators, intravital imaging system is a powerful technique to capture such dynamic responses. We here describe how to evaluate vascular permeability in vivo using multiphoton microscopy. We use various sizes of fluorescence-labeled dextran to visualize how leaky the blood vessels are in the steady state and in inflammation. Using this assay system, we can illustrate the dynamic kinetics of vascular permeability in vivo in real-time. This assay system provides a novel convenient way to study vascular biology that is beneficial in the assessment of various animal models of allergic disease.

Multimodal imaging with spectral-domain optical coherence tomography and photoacoustic remote sensing microscopy.

We develop a multimodal imaging platform, combining depth-resolved scattering contrast from spectral-domain optical coherence tomography (SD-OCT) with complementary, non-contact absorption contrast using photoacoustic remote sensing (PARS) microscopy. The system provides a widefield OCT mode using a telecentric scan lens, and a high-resolution, dual-contrast mode using a 0.26 numerical aperture apochromatic objective. An interlaced acquisition approach is used to achieve simultaneous, co-registered imaging. The SD-OCT modality provides a 9.7 µm axial resolution. Comprehensive in vivo imaging of a nude mouse ear is demonstrated, with the SD-OCT scattering intensity revealing dermal morphology, and PARS microscopy providing a map of microvasculature.

High-resolution computed tomography evaluation of congenital aural atresia - how useful is this?

OBJECTIVE: To depict various temporal bone abnormalities on high-resolution computed tomography in congenital aural atresia patients, and correlate these findings with auditory function test results and microtia subgroup. METHODS: Forty patients (56 ears) with congenital malformation of the auricle and/or external auditory canal were evaluated. Auricles were graded according to Marx's classification, divided into subgroups of minor (grades I and II) and major (III and IV) microtia. Other associated anomalies of the external auditory canal, tympanic cavity, ossicular status, oval and round windows, facial nerve, and inner ear were evaluated. RESULTS: Minor and major microtia were observed in 53.6 and 46.4 per cent of ears respectively. Mean hearing levels were 62.47 and 62.37 dB respectively (p = 0.98). The malleus was the most commonly dysplastic ossicle (73.3 vs 80.8 per cent of ears respectively, p = 0.53). Facial nerve (mastoid segment) abnormalities were associated (p = 0.04) with microtia subgroup (80 vs 100 per cent in minor vs major subgroups). CONCLUSION: Microtia grade was not significantly associated with mean hearing levels or other ear malformations, except for external auditory canal and facial nerve (mastoid segment) anomalies. High-resolution computed tomography is essential in congenital aural atresia, before management strategy is decided.

Is There a Coexistence of Peritemporal Wormian Bones and Congenital Aural Atresia?

In this study, the percent of wormian bones in congenital aural atresia group was higher than healthy group, when tested by the Fisher exact test, there appeared to be no significant statistical difference. However, in congenital aural atresia surgery, surgical manipulation can dislodge these bones causing injury to the surrounding structures. Therefore, the radiologists should report the presence of wormian bones in the peritemporal area.

Methods of Estimating Nasogastric Tube Length: All, Including "NEX," Are Unsafe.

BACKGROUND: Predominance of blind feeding tube placement makes esophageal tube misplacement and aspiration risk commonplace. Accurate estimation of nose-to-stomach length could reduce this risk. Standards for estimating this length were audited against the length measured from guided tube placement. METHODS: This prospective, single-center observational study used electromagnet-guided tube placement to measure the length from nose to gastric body flexure as part of routine care. This measurement was used to audit standard equations used to estimate this length from external measures: xiphisternum-ear-nose + 10 cm (XEN+10), nose-ear-xiphisternum (NEX), and Hanson_A and Hanson_B. RESULTS: From April 23, 2015, to March 2, 2020, measurements were obtained from 200 primary tube placements. Median length to the gastric body flexure (61 cm) was significantly different from that to the pre-gastroesophageal junction flexure (48 cm) or lengths predicted by NEX (51 cm) or Hanson_A (50.5 cm) and Hanson_B (56.1 cm) (all P < .00001) but similar to XEN+10 (61 cm). Esophageal placement was a potential risk for all methods (NEX: 96.3%, Hanson_A: 99.5%, Hanson_B: 86.9%, XEN+10: 43.2%) and a definite risk for most (NEX and Hanson_A: 14.9%, Hanson_B: 1%, XEN+10: 0%). CONCLUSIONS: NEX and Hanson methods of predicting the length from nose to gastric body flexure are too short and risk esophageal misplacement. XEN+10 reduces but does not eliminate this risk. External measurement predictions are clinically unsafe as a guide blind tube placement. Guided placement is recommended.

Transparent High-Frequency Ultrasonic Transducer for Photoacoustic Microscopy Application.

We report the development of an optically transparent high-frequency ultrasonic transducer using lithium niobate single-crystal and indium-tin-oxide electrodes with up to 90% optical transmission in the visible-to-near-infrared spectrum. The center frequency of the transducer was at 36.9 MHz with 33.9%, at -6 dB fractional bandwidth. The photoacoustic imaging capability of the fabricated transducer was also demonstrated by successfully imaging a resolution target and mouse-ear vasculatures in vivo, which were irradiated by a 532 nm pulse laser transmitted through the transducer.

Triplex radiometric, photoacoustic, and ultrasonic imaging based on single-pulse excitation.

In this Letter, we propose a novel triplex-parameter detection method to realize simultaneous radiometric, photoacoustic, and ultrasonic imaging based on single-pulse excitation. The optical attenuation, optical absorption, and acoustic impedance properties can be obtained simultaneously by analyzing the photoacoustic signals and the ultrasonic echo signals. To test the feasibility and accuracy of this method, agar phantoms with different absorption coefficients and elastic coefficients were measured. Then, this method was experimentally verified by imaging a leaf skeleton piece embedded in an agar cylinder. Furthermore, pilot experiments were performed by triplex imaging of pig ear tissue ex vivo to characterize the cartilage and surrounding tissue. Experimental results demonstrated that this technique has future potentials for visualizing and providing the functional and structural information of biological tissues.

Location of the stapedius muscle with reference to the facial nerve in patients with unilateral congenital aural atresia: implication for active middle ear implants surgery.

Background: Detailed investigations of the stapedial muscle (SM) in congenital aural atresia (CAA) patients have yet to be adequately conducted.Objectives: To assess image variations in the mastoid segment of the facial nerve (FN) and SM in CAA.Materials and methods: A total of nine patients comprising of 9 ears with unilateral CAA were studied. The courses of the FN and SM were evaluated from the basic point to 1 mm intervals between the mastoid portion of FN, and measured from the mean X and Y values in each group.Results: The atresia side of FN among the Y values showed significant differences compared to the contralateral side. In terms of the SM, there were no significant differences in both the X and Y values. The stapedial muscle of the CAA patients was located medially to the FN. Conversely, the distance from the PSC to the FN revealed no significant differences with regard to the X and Y values for each group.Conclusion: The current observations revealed that the SM is located more posterior to the FN in CAA patients, and this is mainly attributed to the laterally and anteriorly displaced FN.