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Sample preparation is a crucial step in single-molecule experiments and involves passivating the microfluidic sample chamber, immobilizing the molecules, and setting experimental buffer conditions. The efficiency of the experiment depends on the quality and speed of sample preparation, which is often performed manually and relies on the experience of the experimenter. This can result in inefficient use of single-molecule samples and time, especially for high-throughput applications. To address this, a pressure-controlled microfluidic system is proposed to automate single-molecule sample preparation. The hardware is based on microfluidic components from ElveFlow and is designed to be cost-effective and adaptable to various microscopy applications. The system includes a reservoir pressure adapter and a reservoir holder designed for additive manufacturing. Two flow chamber designs Ibidi µ-slide and Grace Bio-Labs HybriWell chamber are characterized, and the flow characteristics of the liquid at different volume flow rates VË are simulated using CFD-simulations and compared to experimental and theoretical values. The goal of this work is to establish a straightforward and robust system for single-molecule sample preparation that can increase the efficiency of experiments and reduce the bottleneck of manual sample preparation, particularly for high-throughput applications.
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The human voice signal originates from the vibrations of the two vocal folds within the larynx. The interactions of several intrinsic laryngeal muscles adduct and shape the vocal folds to facilitate vibration in response to airflow. Three-dimensional vocal fold dynamics are extracted from in vitro hemilarynx experiments and fitted by a numerical three-dimensional-multi-mass-model (3DM) using an optimization procedure. In this work, the 3DM dynamics are optimized over 24 experimental data sets to estimate biomechanical vocal fold properties during phonation. Accuracy of the optimization is verified by low normalized error (0.13 ± 0.02), high correlation (83% ± 2%), and reproducible subglottal pressure values. The optimized, 3DM parameters yielded biomechanical variations in tissue properties along the vocal fold surface, including variations in both the local mass and stiffness of vocal folds. That is, both mass and stiffness increased along the superior-to-inferior direction. These variations were statistically analyzed under different experimental conditions (e.g., an increase in tension as a function of vocal fold elongation and an increase in stiffness and a decrease in mass as a function of glottal airflow). The study showed that physiologically relevant vocal fold tissue properties, which cannot be directly measured during in vivo human phonation, can be captured using this 3D-modeling technique.
Assuntos
Fonação/fisiologia , Prega Vocal/fisiologia , Idoso , Fenômenos Biomecânicos/fisiologia , Cadáver , Elasticidade , Humanos , Masculino , Modelos Biológicos , Pressão , Prega Vocal/anatomia & histologiaRESUMO
With the use of an endoscopic, high-speed camera, vocal fold dynamics may be observed clinically during phonation. However, observation and subjective judgment alone may be insufficient for clinical diagnosis and documentation of improved vocal function, especially when the laryngeal disease lacks any clear morphological presentation. In this study, biomechanical parameters of the vocal folds are computed by adjusting the corresponding parameters of a three-dimensional model until the dynamics of both systems are similar. First, a mathematical optimization method is presented. Next, model parameters (such as pressure, tension and masses) are adjusted to reproduce vocal fold dynamics, and the deduced parameters are physiologically interpreted. Various combinations of global and local optimization techniques are attempted. Evaluation of the optimization procedure is performed using 50 synthetically generated data sets. The results show sufficient reliability, including 0.07 normalized error, 96% correlation, and 91% accuracy. The technique is also demonstrated on data from human hemilarynx experiments, in which a low normalized error (0.16) and high correlation (84%) values were achieved. In the future, this technique may be applied to clinical high-speed images, yielding objective measures with which to document improved vocal function of patients with voice disorders.
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Simulação por Computador , Modelos Biológicos , Fonação , Prega Vocal/fisiologia , Voz , Algoritmos , Fenômenos Biomecânicos , Feminino , Humanos , Laringoscopia , Masculino , Pressão , Reprodutibilidade dos Testes , Vibração , Gravação em Vídeo , Prega Vocal/anatomia & histologiaRESUMO
In this work a detection algorithm for mucosal wave propagation is presented. By incorporating physiological knowledge of mucosal wave properties and taking the segmented lateral movement of both vocal fold edges as a basis, the spatio-temporal position of the traveling mucosal wave is identified and quantitatively captured. The course of mucosal wave propagation can be successfully detected and analyzed with regard to discriminating different types of mucosal wave activity (in terms of spread velocity and symmetry). The preliminary results obtained for six exemplary laryngeal high-speed recordings are promising and demonstrate the potential of the proposed detection and objective description approach.
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Mucosa Laríngea/fisiologia , Modelos Biológicos , Acústica da Fala , Prega Vocal/fisiologia , Voz/fisiologia , Algoritmos , HumanosRESUMO
Human voice originates from the three-dimensional (3D) oscillations of the vocal folds. In previous studies, biomechanical properties of vocal fold tissues have been predicted by optimizing the parameters of simple two-mass-models to fit its dynamics to the high-speed imaging data from the clinic. However, only lateral and longitudinal displacements of the vocal folds were considered. To extend previous studies, a 3D mass-spring, cover-model is developed, which predicts the 3D vibrations of the entire medial surface of the vocal fold. The model consists of five mass planes arranged in vertical direction. Each plane contains five longitudinal, mass-spring, coupled oscillators. Feasibility of the model is assessed using a large body of dynamical data previously obtained from excised human larynx experiments, in vivo canine larynx experiments, physical models, and numerical models. Typical model output was found to be similar to existing findings. The resulting model enables visualization of the 3D dynamics of the human vocal folds during phonation for both symmetric and asymmetric vibrations.
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Modelos Biológicos , Prega Vocal/fisiologia , Pressão do Ar , Algoritmos , Animais , Fenômenos Biomecânicos , Simulação por Computador , Bases de Dados Factuais , Cães , Estudos de Viabilidade , Glote/fisiologia , Humanos , Técnicas In Vitro , Laringe/fisiologia , Periodicidade , Fonação/fisiologia , VibraçãoRESUMO
The clinical diagnosis of voice disorders is based on examination of the rapidly moving vocal folds during phonation (f0: 80-300Hz) with state-of-the-art endoscopic high-speed cameras. Commonly, analysis is performed in a subjective and time-consuming manner via slow-motion video playback and exhibits low inter- and intra-rater reliability. In this study an objective method to overcome this drawback is presented being based on Phonovibrography, a novel image analysis technique. For a collective of 45 normophonic and paralytic voices the laryngeal dynamics were captured by specialized Phonovibrogram features and analyzed with different machine learning algorithms. Classification accuracies reached 93% for 2-class and 73% for 3-class discrimination. The results were validated by subjective expert ratings given the same diagnostic criteria. The automatic Phonovibrogram analysis approach exceeded the experienced raters' classifications by 9%. The presented method holds a lot of potential for providing reliable vocal fold diagnosis support in the future.
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Inteligência Artificial , Diagnóstico por Computador , Laringoscopia/instrumentação , Paralisia das Pregas Vocais/diagnóstico , Prega Vocal/patologia , Adulto , Algoritmos , Simulação por Computador , Interpretação Estatística de Dados , Feminino , Humanos , Interpretação de Imagem Assistida por Computador/instrumentação , Interpretação de Imagem Assistida por Computador/métodos , Reconhecimento Automatizado de Padrão , Fonação , Gravação em Vídeo/instrumentação , Gravação em Vídeo/métodos , Paralisia das Pregas Vocais/patologiaRESUMO
OBJECTIVE: This work presents a computer-aided method for automatically and objectively classifying individuals with healthy and dysfunctional vocal fold vibration patterns as depicted in clinical high-speed (HS) videos of the larynx. METHODS: By employing a specialized image segmentation and vocal fold movement visualization technique - namely phonovibrography - a novel set of numerical features is derived from laryngeal HS videos capturing the dynamic behavior and the symmetry of oscillating vocal folds. In order to assess the discriminatory power of the features, a support vector machine is applied to the preprocessed data with regard to clinically relevant diagnostic tasks. Finally, the classification performance of the learned nonlinear models is evaluated to allow for conclusions to be drawn about suitability of features and data resulting from different examination paradigms. As a reference, a second feature set is determined which corresponds to more traditional voice analysis approaches. RESULTS: For the first time an automatic classification of healthy and pathological voices could be obtained by analyzing the vibratory patterns of vocal folds using phonovibrograms (PVGs). An average classification accuracy of approximately 81% was achieved for 2-class discrimination with PVG features. This exceeds the results obtained through traditional voice analysis features. Furthermore, a relevant influence of phonation frequency on classification accuracy was substantiated by the clinical HS data. CONCLUSION: The PVG feature extraction and classification approach can be assessed as being promising with regard to the diagnosis of functional voice disorders. The obtained results indicate that an objective analysis of dysfunctional vocal fold vibration can be achieved with considerably high accuracy. Moreover, the PVG classification method holds a lot of potential when it comes to the clinical assessment of voice pathologies in general, as the diagnostic support can be provided to the voice clinician in a timely and reliable manner. Due to the observed interdependency between phonation frequency and classification accuracy, in future comparative studies of HS recordings of oscillating vocal folds homogeneous frequencies should be taken into account during examination.