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.

Miniaturized preamplifier integration in ultrasound transducer design for enhanced photoacoustic imaging.

Photoacoustic imaging (PAI) utilizes the photoacoustic effect to record both vascular and functional characteristics of a biological tissue. Photoacoustic signals have typically low amplitude that cannot be read efficiently by data acquisition systems. This necessitates the use of one or more amplifiers. These amplifiers are somewhat bulky (e.g., the ZFL-500LN+, Mini-Circuits, USA, or 351A-3-50-NI, Analog Modules Inc., USA). Here, we describe the fabrication and development process of a transducer with a built-in low-noise preamplifier that is encased within the transducer housing. This new, to the best of our knowledge, design could be advantageous for applications where a compact transducer + preamplifier is required. We demonstrate the performance of this compact detection unit in a laser scanning photoacoustic microscopy system by imaging a rat ear ex vivo and a rat brain vasculature in vivo.

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.

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.

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.

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.

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.

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.

Ear-EEG-based sleep scoring in epilepsy: A comparison with scalp-EEG.

Ear-EEG is a wearable electroencephalogram-recording device. It relies on recording electrodes that are nested within a custom-fitted earpiece in the external ear canal. The concept has previously been tested for seizure detection in epileptic patients and for sleep recordings in a healthy population. This study is the first to examine the use of ear-EEG recordings for sleep staging in patients with epilepsy, comparing it with standard recordings from scalp-EEG. We use individuals with epilepsy because of their multiple sleep disturbances, and their complex relationship between seizures and sleep, which make this group very likely to benefit from wearable electroencephalogram devices for sleep if it were introduced in the clinic. The accuracy of the ear-EEG against that of the scalp-EEG is compared for sleep staging, and we evaluate features of sleep architecture in individuals with epilepsy. A mean kappa value of 0.74 is found for the agreement between hypnograms derived from ear-EEG and scalp-EEG. Furthermore, it was discovered that sleep stage transition frequency could be contributing to the kappa variation. These findings are related to other ear-recording systems in the literature, and the potentials and future obstacles of the device are discussed.

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.

Depth resolved label-free multimodal optical imaging platform to study morpho-molecular composition of tissue.

Multimodal imaging platforms offer a vast array of tissue information in a single image acquisition by combining complementary imaging techniques. By merging different systems, better tissue characterization can be achieved than is possible by the constituent imaging modalities alone. The combination of optical coherence tomography (OCT) with non-linear optical imaging (NLOI) techniques such as two-photon excited fluorescence (TPEF), second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) provides access to detailed information of tissue structure and molecular composition in a fast, label-free and non-invasive manner. We introduce a multimodal label-free approach for morpho-molecular imaging and spectroscopy and validate the system in mouse skin demonstrating the potential of the system for colocalized acquisition of OCT and NLOI signals.

Posterior Distraction First or Fronto-Orbital Advancement First for Severe Syndromic Craniosynostosis.

PURPOSE: Posterior calvarial vault expansion using distraction osteogenesis is performed for syndromic craniosynostosis as the first choice. This procedure allows far greater intracranial volume than fronto-orbital advancement (FOA). This study aimed to determine the most suitable timing of posterior distraction or FOA to sufficiently increase the intracranial volume and remodel the skull shape. PATIENTS AND METHODS: From 2014 to 2017, the authors performed posterior distraction in 13 patients with syndromic craniosynostosis. Data on premature suture fusion, age at first visit, age at surgery, skull thickness, and complications were collected. RESULTS: Five patients underwent posterior distraction at approximately 12 months of age and had no complications, including cerebrospinal fluid leakage or gull wing deformity. However, during the waiting period for the operation, the skull deformity continues to extend upward (turribrachycephaly). To prevent progress of the skull deformity, the authors performed the operation at approximately 6 months of age in 7 patients. However, in 3 of 7 patients whose lambdoid sutures were opening, gull wing deformity occurred. From these results, in a patient with severe Beare-Stevenson syndrome, the authors performed FOA first at 5 months of age, followed by posterior distraction at 12 months of age, and achieved favorable results. CONCLUSIONS: Treatment patterns are patient specific and should be tailored to premature suture fusion, specific skull deformity, and required intracranial volume of each patient.

Dual-Modal In Vivo Fluorescence/Photoacoustic Microscopy Imaging of Inflammation Induced by GFP-Expressing Bacteria.

In this study, dual-modal fluorescence and photoacoustic microscopy was performed for noninvasive and functional in vivo imaging of inflammation induced by green fluorescent protein (GFP) transfected bacteria in mice ear. Our imaging results demonstrated that the multimodal imaging technique is able to monitor the tissue immunovascular responses to infections with molecular specificity. Our study also indicated that the combination of photoacoustic and fluorescence microscopy imaging can simultaneously track the biochemical changes including the bacterial distribution and morphological change of blood vessels in the biological tissues with high resolution and enhanced sensitivity. Consequently, the developed method paves a new avenue for improving the understanding of the pathology mechanism of inflammation.

The location of the fetal ears: A hint for prenatal diagnosis of agnathia-otocephaly complex.

Otocephaly is an extremely rare lethal congenital anomaly characterized by the absence or underdevelopment of the mandible. The clinical presentation is variable. Some cases may present with severe micrognathia as the only anomaly seen prenatally. The key to early diagnosis is careful assessment of the location of the fetal ears on 2D ultrasound examination.

Clinical techniques in Veterinary Dermatology: Regional anaesthesia of the canine ear.

BACKGROUND: Video otoscopy is a valuable tool for the diagnosis and treatment of otitis externa (OE) and otitis media (OM) in dogs. Procedures performed by video otoscopy often are painful and it can be challenging to maintain steady anaesthetic depth and provide smooth anaesthetic recovery. A multimodal approach to analgesia and anaesthesia is ideal to minimize complications associated with high doses of anaesthetic drugs while providing the patient with adequate analgesia. HYPOTHESIS/OBJECTIVE: To describe a technique for regional anaesthesia of the canine ear. ANIMALS: One canine cadaver, obtained as part of a teaching laboratory class for other clinical techniques, with institutional approval, was used for illustration purposes. RESULTS: Clinical technique for regional anaesthesia of the canine ear. CONCLUSIONS AND CLINICAL IMPORTANCE: Nerve blocks for regional anaesthesia are an important aspect of a multimodal anaesthetic approach. A technique for blocking the two major sensory nerves to the canine ear is described.

Proposal of a Budget-Friendly Camera Holder for Endoscopic Ear Surgery.

Endoscopic ear surgery (EES) is increasingly a preferred technique in otologic society. It offers excellent visualization of the anatomical structures directly and behind the corners with variable angled telescopes. It also provides reduced operative morbidity due to being able to perform surgical interventions with less invasive approaches. Operative preparation and setup time and cost of endoscopy system are less expensive compared with surgical microscopes. On the other hand, the main disadvantage of EES is that the surgery has to be performed with 1 single hand. It is certainly restrictive for an ear surgeon who has been operating with 2 hands under otologic microscopic views for years and certainly requires a learning period and perseverance. Holding the endoscope by a second surgeon is not executable because of insufficient surgical space.Endoscope/camera holders have been developed for those who need the comfort and convenience afforded by double-handed microscopic ear surgery. An ideal endoscope holder should be easy-to-set up, easily controlled, providing a variety of angled views, allowing the surgeon to operate with 2 hands and, budget-friendly. In this article, a commercially available 11-inch magic arm camera holder is proposed by the authors to be used in EES due to its versatile, convenient, and budget-friendly features. It allows 2-handed EES through existing technology and is affordable for surgeons looking for a low-cost and practical solution.

Anthropometric analysis of 3D ear scans of Koreans and Caucasians for ear product design.

The present study measured 25 dimensions of the ear including the concha and ear canal for ergonomic design of ear products and compared with existing ear measurement studies. Scanning and casting methods were employed to produce 3D ear images for 230 Koreans and 96 Caucasians and measurements of the ear dimensions were obtained by identifying 21 landmarks on individual ear scan image. The Korean ear measurements were found significantly larger (mean difference d¯ = 0.4-3.7 mm) and more varied (ratio of SDs =1.01-1.55) than those of Caucasians in most of ear dimensions. The average ear length and ear breadth of male were significantly longer ( d¯ = 1.3-7.0 mm) and wider ( d¯ = 0.8-3.0 mm) than those of female. Use of gender- and ethnicity-composite ear data is recommended in product design due to the much larger intra-population variations (7.5-22.2 mm) than the corresponding inter-population variations. Practitioner Summary: The 3D ear measurements of Koreans and Caucasians were collected and compared with those of different ethnic populations. The distinct ear features of the populations identified in this study are applicable to ergonomic design of ear products with better fit and comfort. Abbreviations: CCW: cavum concha width; CV: coefficient of variation; EB: ear breadth; EL: ear length; SD: standard deviation; SE: sampling error; 3D: 3 dimensional.

Ultrasonic detection based on polarization-dependent optical reflection.

Owing to their extremely wide bandwidths, pure optical ultrasonic detection methods are gaining increasing interest. In this Letter, we proposed a simple ultrasonic detector that is based on the polarization-dependent optical reflection. When the acoustic wave reaches the liquid-glass interface, the acoustic pressure changed the relative refractive index between two media, leading to perturbations in the reflectance of the optical probe beam in glass. Unlike previous studies that detected the modulations in the intensity of the reflected beam, our method, named "polarization-dependent reflection ultrasonic detection (PRUD)," detects the intensity difference between two polarization components of the same probe beam. The PRUD significantly increased the sensitivity. Besides a phantom study, we also successfully detect weak photoacoustic waves in an in vivo animal experiment. This novel method can provide a simple way for ultrasonic detection, which will have great potential for ultrasound and photoacoustic imaging and sensing.

Photoacoustic endomicroscopy based on a MEMS scanning mirror.

In this Letter, we present a high-resolution photoacoustic endomicroscopy probe based on a microelectromechanical systems (MEMS) scanning mirror. The built-in optical assembly consists of a 0.7 mm graded-index (GRIN) lens for light focusing and a ϕ1 mm MEMS mirror to reflect and scan the beam. A miniaturized unfocused ultrasound transducer with a center frequency of 10 MHz is used for photoacoustic detection. Sharp blades, carbon fibers, and black tapes were utilized to evaluate the performance of the system. In vivo mouse ears and resected rectums were imaged to further demonstrate the feasibility of this probe for potential biological and clinical applications.

Morphological characteristics of external auditory canal in congenital aural stenosis patients.

Objective To investigate characteristics of congenital aural stenosis (CAS) patients' external auditory canal (EAC) (position, length, orientation, etc.) and compare them with normal EAC. METHODS: CT images of normal people and CAS patient were utilized. We obtained coordinates of EAC landmarks. Then the Matlab program could calculate some anatomic parameters about EAC, including distances from central point of tympanic annulus (CA), central point of osseous EAC opening (CO), central point of cartilaginous EAC inside opening (CCi), central point of cartilaginous EAC outside opening (CCo) to the Frankfurt horizontal plane (Pfrkt), the median sagittal plane (Psag), the coronal plane (Pcor); orientations of EAC bendings; straight and arc lengths of EAC. RESULTS: Distances from CA, CO, CCi and CCo to Pfrkt were all shorter in CAS group than control group (p<0.05). The straight and arc lengths of cartilaginous EAC in CAS group were shorter than control group (p<0.05). Straight and arc lengths of EAC in CAS group were shorter than those in control group (p<0.05). The proportion of one bending in cartilaginous EAC in control group was significantly lower than CAS group (p<0.05). Orientations of EAC bendings in CAS group differed from those in control group (p<0.05). CONCLUSION: In addition to smaller diameters, compared with normal EAC, the position of CAS patients' osseous EAC was higher compared with the normal. The majority of CAS patients have a bending and downward slanting cartilaginous EAC. Orientations of EAC bending in CAS patients were different from normal. Besides, the length of CAS patients' cartilaginous EAC was shorter. However, there were no significant differences between CAS patients and normal people in length of osseous EAC. These differences in anatomic parameters could provide the basis for optimizing the meatoplasty.