Effect of functional electric stimulation on phonation in an ex vivo aged ovine model.

With age, the atrophy of the thyroarytenoid muscle (TAM), and thus atrophy of the vocal folds, leads to decreased glottal closure, increased breathiness, and a loss in voice quality, which results in a reduced quality of life. A method to counteract the atrophy of the TAM is to induce hypertrophy in the muscle by functional electric stimulation (FES). In this study, phonation experiments were performed with ex vivo larynges of six stimulated and six unstimulated ten-year-old sheep to investigate the impact of FES on phonation. Electrodes were implanted bilaterally near the cricothyroid joint. FES treatment was provided for nine weeks before harvesting. The multimodal measurement setup simultaneously recorded high-speed video of the vocal fold oscillation, the supraglottal acoustic signal, and the subglottal pressure signal. Results of 683 measurements show a 65.6% lower glottal gap index, a 22.7% higher tissue flexibility (measured by the amplitude to length ratio), and a 473.7% higher coefficient of determination (R2) of the regression of subglottal and supraglottal cepstral peak prominence during phonation for the stimulated group. These results suggest that FES improves the phonatory process for aged larynges or presbyphonia.

Synthetic mucus for an ex vivo phonation setup: Creation, application, and effect on excised porcine larynges.

Laryngeal mucus hydrates and lubricates the deformable tissue of the vocal folds and acts as a boundary layer with the airflow from the lungs. However, the effects of the mucus' viscoelasticity on phonation remain widely unknown and mucus has not yet been established in experimental procedures of voice research. In this study, four synthetic mucus samples were created on the basis of xanthan with focus on physiological frequency-dependent viscoelastic properties, which cover viscosities and elasticities over 2 orders of magnitude. An established ex vivo experimental setup was expanded by a reproducible and controllable application method of synthetic mucus. The application method and the suitability of the synthetic mucus samples were successfully verified by fluorescence evidence on the vocal folds even after oscillation experiments. Subsequently, the impact of mucus viscoelasticity on the oscillatory dynamics of the vocal folds, the subglottal pressure, and acoustic signal was investigated with 24 porcine larynges (2304 datasets). Despite the large differences of viscoelasticity, the phonatory characteristics remained stable with only minor statistically significant differences. Overall, this study increased the level of realism in the experimental setup for replication of the phonatory process enabling further research on pathological mucus and exploration of therapeutic options.

Fluid-structure-acoustic interactions in an ex vivo porcine phonation model.

In the clinic, many diagnostic and therapeutic procedures focus on the oscillation patterns of the vocal folds (VF). Dynamic characteristics of the VFs, such as symmetry, periodicity, and full glottal closure, are considered essential features for healthy phonation. However, the relevance of these individual factors in the complex interaction between the airflow, laryngeal structures, and the resulting acoustics has not yet been quantified. Sustained phonation was induced in nine excised porcine larynges without vocal tract (supraglottal structures had been removed above the ventricular folds). The multimodal setup was designed to simultaneously control and monitor key aspects of phonation in the three essential parts of the larynx. More specifically, measurements will comprise (1) the subglottal pressure signal, (2) high-speed recordings in the glottal plane, and (3) the acoustic signal in the supraglottal region. The automated setup regulates glottal airflow, asymmetric arytenoid adduction, and the pre-phonatory glottal gap. Statistical analysis revealed a beneficial influence of VF periodicity and glottal closure on the signal quality of the subglottal pressure and the supraglottal acoustics, whereas VF symmetry only had a negligible influence. Strong correlations were found between the subglottal and supraglottal signal quality, with significant improvement of the acoustic quality for high levels of periodicity and glottal closure.

Dynamic Biomechanical Analysis of Vocal Folds Using Pipette Aspiration Technique.

The voice producing process is a complex interplay between glottal pressure, vocal folds, their elasticity and tension. The material properties of vocal folds are still insufficiently studied, because the determination of material properties in soft tissues is often difficult and connected to extensive experimental setups. To shed light on this less researched area, in this work, a dynamic pipette aspiration technique is utilized to measure the elasticity in a frequency range of 100-1000 Hz. The complex elasticity could be assessed with the phase shift between exciting pressure and tissue movement. The dynamic pipette aspiration setup has been miniaturized with regard to a future in-vivo application. The techniques were applied on 3 different porcine larynges 4 h and 1 d postmortem, in order to investigate the deterioration of the tissue over time and analyze correlation in elasticity values between vocal fold pairs. It was found that vocal fold pairs do have different absolute elasticity values but similar trends. This leads to the assumption that those trends are more important for phonation than having same absolute values.

A Quasi-Static Quantitative Ultrasound Elastography Algorithm Using Optical Flow.

Ultrasound elastography is a constantly developing imaging technique which is capable of displaying the elastic properties of tissue. The measured characteristics could help to refine physiological tissue models, but also indicate pathological changes. Therefore, elastography data give valuable insights into tissue properties. This paper presents an algorithm that measures the spatially resolved Young's modulus of inhomogeneous gelatin phantoms using a CINE sequence of a quasi-static compression and a load cell measuring the compressing force. An optical flow algorithm evaluates the resulting images, the stresses and strains are computed, and, conclusively, the Young's modulus and the Poisson's ratio are calculated. The whole algorithm and its results are evaluated by a performance descriptor, which determines the subsequent calculation and gives the user a trustability index of the modulus estimation. The algorithm shows a good match between the mechanically measured modulus and the elastography result-more precisely, the relative error of the Young's modulus estimation with a maximum error 35%. Therefore, this study presents a new algorithm that is capable of measuring the elastic properties of gelatin specimens in a quantitative way using only the image data. Further, the computation is monitored and evaluated by a performance descriptor, which measures the trustability of the results.

Analysis of the tonal sound generation during phonation with and without glottis closure.

The human phonation is characterized by periodical oscillations of the vocal folds with a complete glottis closure. In contrast, a glottal insufficiency (GI) represents an oscillation without glottis closure resulting in a breathy and weak voice. In this study, flow-induced oscillations of silicone vocal folds were modeled with and without glottis closure. The measurements comprised the flow pressure in the model, the generated sound, and the high-speed footage of the vocal fold motion. The analysis revealed that the sound signal for vocal fold oscillations without closure exhibits a lower number of harmonic tones with smaller amplitudes compared to the case with complete closure. The time series of the pressure signals showed small and periodical oscillations occurring less frequently and with smaller amplitude for the GI case. Accordingly, the pressure spectra include fewer harmonics similar to the sound. The analysis of the high-speed videos indicates that the strength of the pressure oscillations correlates with the divergence angle of the glottal duct during the closing motion. Physiologically, large divergence angles typically occur for a pronounced mucosal wave motion with glottis closure. Thus, the results indicate a correlation between the intensity of the mucosal wave and the development of harmonic tones.

Influence of glottal closure on the phonatory process in ex vivo porcine larynges.

Many cases of disturbed voice signals can be attributed to incomplete glottal closure, vocal fold oscillation asymmetries, and aperiodicity. Often these phenomena occur simultaneously and interact with each other, making a systematic, isolated investigation challenging. Therefore, ex vivo porcine experiments were performed which enable direct control of glottal configurations. Different pre-phonatory glottal gap sizes, adduction levels, and flow rates were adjusted. The resulting glottal closure types were identified in a post-processing step. Finally, the acoustic quality, aerodynamic parameters, and the characteristics of vocal fold oscillation were analyzed in reference to the glottal closure types. Results show that complete glottal closure stabilizes the phonation process indicated through a reduced left-right phase asymmetry, increased amplitude and time periodicity, and an increase in the acoustic quality. Although asymmetry and periodicity parameter variation covers only a small range of absolute values, these small variations have a remarkable influence on the acoustic quality. Due to the fact that these parameters cannot be influenced directly, the authors suggest that the (surgical) reduction of the glottal gap seems to be a promising method to stabilize the phonatory process, which has to be confirmed in future studies.

Experimental generation of amplitude squeezed vector beams.

We present an experimental method for the generation of amplitude squeezed high-order vector beams. The light is modified twice by a spatial light modulator such that the vector beam is created by means of a collinear interferometric technique. A major advantage of this approach is that it avoids systematic losses, which are detrimental as they cause decoherence in continuous-variable quantum systems. The utilisation of a spatial light modulator (SLM) gives the flexibility to switch between arbitrary mode orders. The conversion efficiency with our setup is only limited by the efficiency of the SLM. We show the experimental generation of Laguerre-Gauss (LG) modes with radial indices 0 or 1 and azimuthal indices up to 3 with complex polarization structures and a quantum noise reduction up to -0.9dB±0.1dB. The corresponding polarization structures are studied in detail by measuring the spatial distribution of the Stokes parameters.

Single-mode squeezing in arbitrary spatial modes.

As the generation of squeezed states of light has become a standard technique in laboratories, attention is increasingly directed towards adapting the optical parameters of squeezed beams to the specific requirements of individual applications. It is known that imaging, metrology, and quantum information may benefit from using squeezed light with a tailored transverse spatial mode. However, experiments have so far been limited to generating only a few squeezed spatial modes within a given setup. Here, we present the generation of single-mode squeezing in Laguerre-Gauss and Bessel-Gauss modes, as well as an arbitrary intensity pattern, all from a single setup using a spatial light modulator (SLM). The degree of squeezing obtained is limited mainly by the initial squeezing and diffractive losses introduced by the SLM, while no excess noise from the SLM is detectable at the measured sideband. The experiment illustrates the single-mode concept in quantum optics and demonstrates the viability of current SLMs as flexible tools for the spatial reshaping of squeezed light.

What kind of phonation causes the strongest vocal fold collision? - A hemi-larynx phonation contact pressure study.

This paper presents a measurement setup which is able to measure the distribution of small scale pressure on an area of 15.2 mm × 30.4 mm with a sample rate up to 1.2 kHz. It was used to investigate the contact pressures of vocal folds during phonation. This was performed in ex vivo experiments of 11 porcine larynges. The contact pressure at the medial surface and other phonation parameters, as the glottal resistance and the closing velocity of the vocal fold, were measured at different adduction and elongation levels and air flow rates. A statistical analysis was carried out. It could be shown that the contact pressure rises, when the vocal fold is manipulated or when the flow rate is increased.

Analyzing Vocal Fold Frequency Dynamics Using High-Speed 3D Laser Video Endoscopy.

OBJECTIVE: To examine changes in lateral and vertical vibratory motion along the anterior, middle, and posterior sections of the vocal folds, as a function of vocal frequency variations. METHODS: Absolute measurements of vocal fold surface dynamics from high-speed videoendoscopy with custom laser endoscope were made on 23 vocally healthy adults during sustained /i:/ production at 10%, 20%, and 80% of pitch range. The 3D parameters of amplitude (mm), maximum velocity opening/closing (mm/s), and mean velocity opening/closing (mm/s) were computed for the lateral and vertical vibratory motion along the anterior, middle, and posterior sections of the vocal folds. Linear mixed model analysis was conducted to evaluate the differences in (a) vocal frequency levels (high vs. normal vs. low pitch), (b) axis level (vertical vs. lateral), (c) position level (anterior vs. middle vs. posterior), and (d) gender differences (male vs. female). RESULTS: Overall, the superior surface vertical motion of the vocal fold is greater compared with the lateral motion, especially in males. Along the superior surface, the mean and maximum closing velocities are greater posteriorly for low pitch. The location (anterior, middle, and posterior) along the superior surface is relevant only for vocal fold closing rather than opening, as the dynamics are different along the various locations. CONCLUSIONS: The study highlights the significance of assessing the vertical motion of the superior surface of the vocal fold to understand the complex dynamics of voice production. LEVEL OF EVIDENCE: NA Laryngoscope, 134:3267-3276, 2024.

Influence of Perspective Distortion in Laryngoscopy.

OBJECTIVE: An experiment with controllable boundaries was designed to assess the influence of the recording angle and distance on two-dimensional (2D) imaging in laryngoscopy and resulting 2D parameter calculation derived from the glottal area waveform (GAW). METHOD: Two high-speed camera setups were used to synchronously record an oscillating synthetic vocal fold (VF) model, simulating a high-speed videoendoscopy. One camera recorded at variable lateral recording angles and a reference camera in superior perspective. This was performed at different physiological recording distances and for two oscillation modes (with/without contacting VFs). The GAW was derived from the segmented glottis, and two parameters each for the categories of symmetry, periodicity, and closure were calculated, as well as two derivative measures. The percentage difference between the variable and reference camera value pairs was calculated, and the angle and height dependencies were quantified using linear regression. RESULTS: The visual perception of a laryngoscopy was found to be influenced by the lateral recording angle, which may lead to misinterpretation of VF symmetry among inexperienced observers. The strongest influence of recording angle was observed for symmetry parameters, the strongest being the Amplitude Symmetry Index with up to 2.6%/° (p < .05). A dependence on the recording distance was only found for the Maximum Area Declination Rate. CONCLUSIONS: The recording angle in 2D laryngoscopy should be carefully considered during visual inspection of the VF dynamics. Most of the investigated objective parameters were unaffected by the examined perspective distortion. However, especially left-right symmetry measures should only be used under controlled boundary conditions to avoid misdiagnosis and misinterpretation. SUPPLEMENTAL MATERIAL: https://doi.org/10.23641/asha.23961183.

Influence of Reduced Saliva Production on Phonation in Patients With Ectodermal Dysplasia.

OBJECTIVE: Patients with ectodermal dysplasia (ED) suffer from an inherited disorder in the development of the ectodermal structures. Besides the main symptoms, i.e. significantly reduced formation/expression of teeth, hair and sweat glands, a decreased saliva production is objectively accounted. In addition to difficulties with chewing/swallowing, ED patients frequently report on the subjective impression of rough and hoarse voices. A correlation between the reduced production of saliva and an affliction of the voice has not yet been investigated objectively for this rare disease. METHODS: Following an established measurement protocol, a study has been conducted on 31 patients with ED and 47 controls (no ED, healthy voice). Additionally, the vocal fold oscillations were recorded by high-speed videoendoscopy (HSV@4 kHz). The glottal area waveform was determined by segmentation and objective glottal dynamic parameters were calculated. The generated acoustic signal was evaluated by objective and subjective measures. The individual impairment was documented by a standardized questionnaire (VHI). Additionally, the amount of generated saliva was measured for a defined period of time. RESULTS: ED patients displayed a significantly reduced saliva production compared to the control group. Furthermore, the auditory-perceptual evaluation yielded significantly higher ratings for breathiness and hoarseness in the voices of male ED patients compared to male controls. The majority of male ED patients (67%) indicated at least minor impairment in the self-evaluation. Objective acoustic measures like Jitter and Shimmer confirmed the decreased acoustic quality in male ED patients, whereas none of the investigated HSV parameters showed significant differences between the test groups. Statistical analysis did not confirm a statistically significant correlation between reduced voice quality and amount of saliva. CONCLUSIONS: An objective impairment of the acoustic outcome was demonstrated for male ED patients. However, the vocal folds dynamics in HSV recordings seem unaffected.

Mechanical Parameters Based on High-Speed Videoendoscopy of the Vocal Folds in Patients With Ectodermal Dysplasia.

OBJECTIVE: Patients suffering from ectodermal dysplasia (ED), which is an inherited disorder in the development of the ectodermal structures, have a significantly reduced expression of teeth, hair, sweat glands, and salivary glands in the respiratory tract including the larynx. Previous studies within the framework of the present project showed a significantly reduced saliva production and an impairment of the acoustic outcome in ED patients compared to the control group. However, until now, no statistically significant difference between EDs and controls could be found regarding vocal fold dynamics in the high-speed videoendoscopy (HSV) recordings using representative parameters on closure, symmetry, and periodicity. The aim of this study is to examine the role of tissue characteristics by means of objective mechanical parameters derived from HSV recordings. METHODS: This study includes 28 ED patients and 42 controls (no ED, healthy voice). The vocal fold oscillations were recorded by high-speed videoendoscopy (HSV@4kHz). Based on the dynamical measures of the glottal area waveform (GAW), objective glottal dynamic parameters associated with tissue properties like flexibility and stiffness were computed. RESULTS: The present evaluation displays a significant difference between male ED patients and male controls concerning the HSV-based mechanical parameters indicating reduced stiffness and increased deformability for the vocal folds of male ED patients. In contrast to strongly amplitude-dependent parameters, the primarily velocity-based parameters showed no statistically significant deviation. CONCLUSIONS: The presented data provides the first promising indication toward the underlying causes on the laryngeal level leading to the voice conspicuities in ED patients. The significant difference concerning the mechanical parameters suggests a different composition of the extracellular matrix of the tissue of the vocal folds of ED patients compared to controls.

Behind the Complex Interplay of Phonation: Investigating Elasticity of Vocal Folds With Pipette Aspiration Technique During Ex Vivo Phonation Experiments.

OBJECTIVES: The vibration of the vocal folds produces the primary sound for the human speech. The vibration depends mainly on the pressure, airflow of the lungs, and the material properties of the vocal folds. In order to change them, muscles in the larynx stretch the vocal folds. This interplay is rarely investigated, but can give insight in the complex process of speech production. Most material properties studies are damaging the tissue; therefore, a nondestructive one is desired. METHODS: An ex vivo phonation experiment combined with the dynamic Pipette Aspiration Technique is used to investigate 10 porcine larynges, under manipulations of different adduction and elongation levels. For each manipulation, the near surface material properties of the vocal folds are measured as well as different phonation parameters like the subglottal pressure, glottal resistance, frequency, and stiffness. Thereby, a high-speed camera was used to record the vocal fold movement. RESULTS: On most of the measured parameters, the manipulations do show an effect. Both manipulations lead to a higher phonation frequency and an increase of the stiffness of the tissue. Comparing both manipulations, the elongation results in higher elasticity values than the adduction. Different measurement parameters have been compared with each other and correlations could be found. Where the strongest correlation are found among the elasticity values of different frequencies. But it can also be seen that the elasticity values correlate with phonation parameters. CONCLUSION: It was possible to produce a data set of 560 measurements in total. To our knowledge, this is the first time Pipette Aspiration Technique was combined with ex vivo phonation measurements for combined measurements. The amount of measurement data made it possible to carry out statistic investigations. The effect of the manipulations on material properties as well as on phonation parameters could be measured and different correlations could be found. The results lead to the hypothesis that the stretch does not have a huge effect on the material properties of the lamina propria, but more on the underlying muscle.

Extent and Effect of Covering Laryngeal Structures with Synthetic Laryngeal Mucus via Two Different Administration Techniques.

OBJECTIVE: The first goal of this study was to investigate the coverage of laryngeal structures using two potential administration techniques for synthetic mucus: inhalation and lozenge ingestion. As a second research question, the study investigated the potential effects of these techniques on standardized voice assessment parameters. METHODS: Fluorescein was added to throat lozenges and to an inhalation solution to visualize the coverage of laryngeal structures through blue light imaging. The study included 70 vocally healthy subjects. Fifty subjects underwent administration via lozenge ingestion and 20 subjects performed the inhalation process. For the first research question, the recordings from the blue light imaging system were categorized to compare the extent of coverage on individual laryngeal structures objectively. Secondly, a standardized voice evaluation protocol was performed before and after each administration to determine any measurable effects of typical voice parameters. RESULTS: The administration via inhalation demonstrated complete coverage of all laryngeal structures, including the vocal folds, ventricular folds, and arytenoid cartilages, as visualized by the fluorescent dye. In contrast, the application of the lozenge predominantly covered the pharynx and laryngeal surface toward the aryepiglottic fold, but not the inferior structures. All in all, the comparison before and after administration showed no clear effect, although a minor deterioration of the acoustic signal was noted in the shimmer and cepstral peak prominence after the inhalation. CONCLUSIONS: Our findings indicate that the inhalation process is a more effective technique for covering deeper laryngeal structures such as the vocal folds and ventricular folds with synthetic mucus. This knowledge enables further in vivo studies on the role of laryngeal mucus in phonation in general, and how it can be substituted or supplemented for patients with reduced glandular activity as well as for heavy voice users.

Long-term performance assessment of fully automatic biomedical glottis segmentation at the point of care.

Deep Learning has a large impact on medical image analysis and lately has been adopted for clinical use at the point of care. However, there is only a small number of reports of long-term studies that show the performance of deep neural networks (DNNs) in such an environment. In this study, we measured the long-term performance of a clinically optimized DNN for laryngeal glottis segmentation. We have collected the video footage for two years from an AI-powered laryngeal high-speed videoendoscopy imaging system and found that the footage image quality is stable across time. Next, we determined the DNN segmentation performance on lossy and lossless compressed data revealing that only 9% of recordings contain segmentation artifacts. We found that lossy and lossless compression is on par for glottis segmentation, however, lossless compression provides significantly superior image quality. Lastly, we employed continual learning strategies to continuously incorporate new data into the DNN to remove the aforementioned segmentation artifacts. With modest manual intervention, we were able to largely alleviate these segmentation artifacts by up to 81%. We believe that our suggested deep learning-enhanced laryngeal imaging platform consistently provides clinically sound results, and together with our proposed continual learning scheme will have a long-lasting impact on the future of laryngeal imaging.

Quasi-static ultrasound elastography of ex-vivo porcine vocal folds during passive elongation and adduction.

OBJECTIVES: The elastic properties of the vocal folds have great influence on the primary sound and thus on the entire subsequent phonation process. Muscle contractions in the larynx can alter the elastic properties of the vocal fold tissue. Quasi-static ultrasound elastography is a non-destructive examination method that can be applied to ex-vivo vocal folds. In this work, porcine vocal folds were passively elongated and adducted and the changes of the elastic properties due to that manipulations were measured. METHODS: Manipulations were performed by applying force to sewn-in sutures. Elongation was achieved by a suture attached to the thyroid cartilage, which was pulled forward by defined weights. Adduction was effected by two sutures exerting torque on the arytenoid cartilage. A series of ten specimens was examined and evaluated using a quasi-static elastography algorithm. In addition, the surface stretch was measured optically using tattooed reference points. RESULTS: This study showed that the expected stiffening of the tissue during the manipulations can be measured using quasi-static ultrasound elastography. The measured effect of elongation and adduction, both of which result in stretching of the tissue, is stiffening. However, the relative change of specific manipulations is not the same for the same load on different larynges, but is rather related to stretch caused and other uninvestigated factors. CONCLUSION: The passive elongation and adduction of vocal folds stiffen the tissue of the vocal folds and can be measured using ultrasound elastography.

A single latent channel is sufficient for biomedical glottis segmentation.

Glottis segmentation is a crucial step to quantify endoscopic footage in laryngeal high-speed videoendoscopy. Recent advances in deep neural networks for glottis segmentation allow for a fully automatic workflow. However, exact knowledge of integral parts of these deep segmentation networks remains unknown, and understanding the inner workings is crucial for acceptance in clinical practice. Here, we show that a single latent channel as a bottleneck layer is sufficient for glottal area segmentation using systematic ablations. We further demonstrate that the latent space is an abstraction of the glottal area segmentation relying on three spatially defined pixel subtypes allowing for a transparent interpretation. We further provide evidence that the latent space is highly correlated with the glottal area waveform, can be encoded with four bits, and decoded using lean decoders while maintaining a high reconstruction accuracy. Our findings suggest that glottis segmentation is a task that can be highly optimized to gain very efficient and explainable deep neural networks, important for application in the clinic. In the future, we believe that online deep learning-assisted monitoring is a game-changer in laryngeal examinations.

Re-Training of Convolutional Neural Networks for Glottis Segmentation in Endoscopic High-Speed Videos.

Endoscopic high-speed video (HSV) systems for visualization and assessment of vocal fold dynamics in the larynx are diverse and technically advancing. To consider resulting "concepts shifts" for neural network (NN)-based image processing, re-training of already trained and used NNs is necessary to allow for sufficiently accurate image processing for new recording modalities. We propose and discuss several re-training approaches for convolutional neural networks (CNN) being used for HSV image segmentation. Our baseline CNN was trained on the BAGLS data set (58,750 images). The new BAGLS-RT data set consists of additional 21,050 images from previously unused HSV systems, light sources, and different spatial resolutions. Results showed that increasing data diversity by means of preprocessing already improves the segmentation accuracy (mIoU + 6.35%). Subsequent re-training further increases segmentation performance (mIoU + 2.81%). For re-training, finetuning with dynamic knowledge distillation showed the most promising results. Data variety for training and additional re-training is a helpful tool to boost HSV image segmentation quality. However, when performing re-training, the phenomenon of catastrophic forgetting should be kept in mind, i.e., adaption to new data while forgetting already learned knowledge.