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This study presents a miniaturized head-mount optical coherence tomography (OCT) system tailored for high-resolution brain imaging in freely moving mice, providing an advanced noninvasive imaging tool in neuroscience research. Leveraging optical coherence tomography technology, the system enables depth-resolved imaging and integrates functional OCT extensions, including angiography and Doppler imaging. Remarkably lightweight at 1.5 g, the device allows for the preservation of natural mouse behavior during imaging sessions. With a maximum 4 × 4 mm field of view and 7.4 µm axial resolution, the system offers reliable imaging capabilities. Noteworthy features include focal adjustability, rotary joint integration for fiber-twist-free operation, and a high-speed swept-source OCT laser at 200 kHz, facilitating real-time imaging. By providing insights into brain mechanisms and neurological disorders without disrupting natural behavior, this innovative system holds promise as a powerful tool in neuroscience research. Its compact design and comprehensive imaging capabilities make it well-suited for studying various brain regions and dynamic processes, contributing significantly to our understanding of brain function and pathology.
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Background: The coordination and the directional order of ciliary metachronal waves are the major factors that determine the effectiveness of mucociliary clearance (MCC). Even though metachronal waves play an essential part in immune response, clinical diagnostic tools and imaging techniques that can reliably and efficiently capture their spatial distribution and function are currently limited. Methods: We present label-free high-speed visualization of ciliary metachronal wave propagations in freshly-excised tracheal explants using a spectrally-encoded interferometric microscope over a two-dimensional (2D) plane of 0.5 mm × 0.5 mm at an acquisition rate of 50 frame-per-second. Furthermore, phase-resolved enhanced dynamic (PHRED) analysis of time-series doppler images was performed, where spatial-temporal characteristics of cilia metachronal wave motions are revealed through frequency component analysis and spatial filtering. Results: The PHRED analysis of phase-resolved Doppler (PRD) images offers a capability to distinguish the propagation direction of metachronal waves, and quantitatively assess amplitude and dominant frequency of cilia beating at each spatial location. Compared to the raw PRD images, the phase-resolved dynamic wavefront imaging (PRDWI) method showed the direction and coordination of collective cilia movement more distinctively. Conclusions: The PRDWI technique can have broad application prospects for the diagnosis of human respiratory diseases and evaluation of the curative effect of treatments and open new perspectives in biomedical sciences.
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Photo-mediated Ultrasound Therapy (PUT), as a new anti-vascular technique, can promote cavitation activity to selectively destruct blood vessels with a significantly lower amount of energy when compared to energy level required by other laser and ultrasound treatment therapies individually. Here, we report the development of a high speed PUT system based on a 50-kHz pulsed laser to achieve faster treatment, decreasing the treatment time by a factor of 20. Furthermore, we integrated it with optical coherence tomography angiography (OCTA) for real time monitoring. The feasibility of the proposed OCTA-guided PUT was validated through in vivo rabbit experiments. The addition of OCTA to PUT allows for quantitative prescreening and real time monitoring of treatment response, thereby enabling implementation of individualized treatment strategies.
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Tomografía de Coherencia Óptica , Terapia por Ultrasonido , Animales , Conejos , Tomografía de Coherencia Óptica/métodos , Angiografía con Fluoresceína/métodos , Terapia por Ultrasonido/métodos , Rayos LáserRESUMEN
SIGNIFICANCE: The human vocal fold (VF) oscillates in multiple vectors and consists of distinct layers with varying viscoelastic properties that contribute to the mucosal wave. Office-based and operative laryngeal endoscopy are limited to diagnostic evaluation of the VF epithelial surface only and are restricted to axial-plane characterization of the horizontal mucosal wave. As such, understanding of the biomechanics of human VF motion remains limited. AIM: Optical coherence tomography (OCT) is a micrometer-resolution, high-speed endoscopic imaging modality which acquires cross-sectional images of tissue. Our study aimed to leverage OCT technology and develop quantitative methods for analyzing the anatomy and kinematics of in vivo VF motion in the coronal plane. APPROACH: A custom handheld laryngeal stage was used to capture OCT images with 800 A-lines at 250 Hz. Automated image postprocessing and analytical methods were developed. RESULTS: Novel kinematic analysis of in vivo, long-range OCT imaging of the vibrating VF in awake human subjects is reported. Cross-sectional, coronal-plane panoramic videos of the larynx during phonation are presented with three-dimensional videokymographic and space-time velocity analysis of VF motion. CONCLUSIONS: Long-range OCT with automated computational methods allows for cross-sectional dynamic laryngeal imaging and has the potential to broaden our understanding of human VF biomechanics and sound production.
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Laringe , Tomografía de Coherencia Óptica , Fenómenos Biomecánicos , Humanos , Fonación , Pliegues Vocales/diagnóstico por imagenRESUMEN
Optical coherence tomography (OCT) has gained wide adoption in biological research and medical imaging due to its exceptional tissue penetration, 3D imaging speed, and rich contrast. However, OCT plays a relatively small role in molecular and cellular imaging due to the lack of suitable biomolecular contrast agents. In particular, while the green fluorescent protein has provided revolutionary capabilities to fluorescence microscopy by connecting it to cellular functions such as gene expression, no equivalent reporter gene is currently available for OCT. Here, we introduce gas vesicles, a class of naturally evolved gas-filled protein nanostructures, as genetically encodable OCT contrast agents. The differential refractive index of their gas compartments relative to surrounding aqueous tissue and their nanoscale motion enables gas vesicles to be detected by static and dynamic OCT. Furthermore, the OCT contrast of gas vesicles can be selectively erased in situ with ultrasound, allowing unambiguous assignment of their location. In addition, gas vesicle clustering modulates their temporal signal, enabling the design of dynamic biosensors. We demonstrate the use of gas vesicles as reporter genes in bacterial colonies and as purified contrast agents in vivo in the mouse retina. Our results expand the utility of OCT to image a wider variety of cellular and molecular processes.
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Nanoestructuras , Tomografía de Coherencia Óptica , Animales , Medios de Contraste , Imagenología Tridimensional , Ratones , UltrasonografíaRESUMEN
BACKGROUND: Acute respiratory distress syndrome (ARDS) is a severe form of acute lung injury with a mortality rate of up to 40%. Early management of ARDS has been difficult due to the lack of sensitive imaging tools and robust analysis software. We previously designed an optical coherence tomography (OCT) system to evaluate mucosa thickness (MT) after smoke inhalation, but the analysis relied on manual segmentation. The aim of this study is to assess in vivo proximal airway volume (PAV) after inhalation injury using automated OCT segmentation and correlate the PAV to lung function for rapid indication of ARDS. METHODS: Anesthetized female Yorkshire pigs (n = 14) received smoke inhalation injury (SII) and 40% total body surface area thermal burns. Measurements of PaO2-to-FiO2 ratio (PFR), peak inspiratory pressure (PIP), dynamic compliance, airway resistance, and OCT bronchoscopy were performed at baseline, postinjury, 24 hours, 48 hours, 72 hours after injury. A tissue segmentation algorithm based on graph theory was used to reconstruct a three-dimensional (3D) model of lower respiratory tract and estimate PAV. Proximal airway volume was correlated with PFR, PIP, compliance, resistance, and MT measurement using a linear regression model. RESULTS: Proximal airway volume decreased after the SII: the group mean of proximal airway volume at baseline, postinjury, 24 hours, 48 hours, 72 hours were 20.86 cm (±1.39 cm), 17.61 cm (±0.99 cm), 14.83 cm (±1.20 cm), 14.88 cm (±1.21 cm), and 13.11 cm (±1.59 cm), respectively. The decrease in the PAV was more prominent in the animals that developed ARDS after 24 hours after the injury. PAV was significantly correlated with PIP (r = 0.48, p < 0.001), compliance (r = 0.55, p < 0.001), resistance (r = 0.35, p < 0.01), MT (r = 0.60, p < 0.001), and PFR (r = 0.34, p < 0.01). CONCLUSION: Optical coherence tomography is a useful tool to quantify changes in MT and PAV after SII and burns, which can be used as predictors of developing ARDS at an early stage. LEVEL OF EVIDENCE: Prognostic, level III.
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Lesión Pulmonar/diagnóstico por imagen , Síndrome de Dificultad Respiratoria/diagnóstico por imagen , Lesión por Inhalación de Humo/diagnóstico por imagen , Tomografía de Coherencia Óptica , Animales , Broncoscopía , Femenino , Lesión Pulmonar/complicaciones , Síndrome de Dificultad Respiratoria/etiología , Lesión por Inhalación de Humo/complicaciones , PorcinosRESUMEN
BACKGROUND: The prevalence of acute respiratory distress syndrome (ARDS) in mechanically ventilated burn patients is 33%, with mortality varying from 11-46% depending on ARDS severity. Despite the new Berlin definition for ARDS, prompt bedside diagnosis is lacking. We developed and tested a bedside technique of fiberoptic-bronchoscopy-based optical coherence tomography (OCT) measurement of airway mucosal thickness (MT) for diagnosis of ARDS following smoke inhalation injury (SII) and burns. METHODS: 16 female Yorkshire pigs received SII and 40% thermal burns. OCT MT and PaO2-to-FiO2 ratio (PFR) measurements were taken at baseline, after injury, and at 24, 48, and 72h after injury. RESULTS: Injury led to thickening of MT which was sustained in animals that developed ARDS. Significant correlations were found between MT, PFR, peak inspiratory pressure (PIP), and total infused fluid volume. CONCLUSIONS: OCT is a useful tool to quantify MT changes in the airway following SII and burns. OCT may be effective as a diagnostic tool in the early stages of SII-induced ARDS and should be tested in humans.
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Broncoscopía/métodos , Síndrome de Dificultad Respiratoria/diagnóstico por imagen , Mucosa Respiratoria/diagnóstico por imagen , Lesión por Inhalación de Humo/diagnóstico por imagen , Tomografía de Coherencia Óptica/métodos , Animales , Quemaduras por Inhalación/complicaciones , Quemaduras por Inhalación/diagnóstico por imagen , Quemaduras por Inhalación/patología , Femenino , Tamaño de los Órganos , Presión Parcial , Pruebas en el Punto de Atención , Síndrome de Dificultad Respiratoria/etiología , Síndrome de Dificultad Respiratoria/patología , Mucosa Respiratoria/patología , Lesión por Inhalación de Humo/complicaciones , Lesión por Inhalación de Humo/patología , Sus scrofa , PorcinosRESUMEN
Ciliary motion plays a critical role in the overall respiratory health of the upper airway. These cilia beat at a native frequency and in a synchronized pattern to continuously transport foreign particulate trapped in a layer of mucous out of the upper airway. Disruption of ciliary motion can lead to severe respiratory diseases and compromised respiratory function. Currently, the study of cilia requires expensive high speed cameras and high powered microscopes which is unsuitable for in vivo imaging and diagnosis. Doppler based optical coherence tomography has the potential to visualize the microscopic motion of cilia during their beating cycle. We demonstrate the development of a high-speed Doppler optical coherence tomography system that not only can rapidly determine the cilia beat frequency, but also simultaneously visualize the temporal cilia beating pattern which plays critical roles in cilia function.
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Cilios/fisiología , Movimiento (Física) , Periodicidad , Mucosa Respiratoria/fisiología , Tomografía de Coherencia Óptica/métodos , Animales , Masculino , Conejos , Tomografía de Coherencia Óptica/instrumentaciónRESUMEN
The upper airway is a complex tissue structure that is prone to collapse. Current methods for studying airway obstruction are inadequate in safety, cost, or availability, such as CT or MRI, or only provide localized qualitative information such as flexible endoscopy. Long range optical coherence tomography (OCT) has been used to visualize the human airway in vivo, however the limited imaging range has prevented full delineation of the various shapes and sizes of the lumen. We present a new long range OCT system that integrates high speed imaging with a real-time position tracker to allow for the acquisition of an accurate 3D anatomical structure in vivo. The new system can achieve an imaging range of 30 mm at a frame rate of 200 Hz. The system is capable of generating a rapid and complete visualization and quantification of the airway, which can then be used in computational simulations to determine obstruction sites.
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Sistema Respiratorio/anatomía & histología , Tomografía de Coherencia Óptica/métodos , Simulación por Computador , Humanos , Imagenología Tridimensional/métodosRESUMEN
Diagnosis and treatment of vocal fold lesions has been a long-evolving science for the otolaryngologist. Contemporary practice requires biopsy of a glottal lesion in the operating room under general anesthesia for diagnosis. Current in-office technology is limited to visualizing the surface of the vocal folds with fiber-optic or rigid endoscopy and using stroboscopic or high-speed video to infer information about submucosal processes. Previous efforts using optical coherence tomography (OCT) have been limited by small working distances and imaging ranges. Here we report the first full field, high-speed, and long-range OCT images of awake patients' vocal folds as well as cross-sectional video and Doppler analysis of their vocal fold motions during phonation. These vertical-cavity surface-emitting laser source (VCSEL) OCT images offer depth resolved, high-resolution, high-speed, and panoramic images of both the true and false vocal folds. This technology has the potential to revolutionize in-office imaging of the larynx.
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Tomografía de Coherencia Óptica/instrumentación , Disfunción de los Pliegues Vocales/diagnóstico por imagen , Humanos , Laringoscopía/instrumentación , Laringoscopía/métodos , Tomografía de Coherencia Óptica/métodos , Pliegues Vocales/diagnóstico por imagenRESUMEN
RATIONALE: Subglottic edema and acquired subglottic stenosis are potentially airway-compromising sequelae in neonates following endotracheal intubation. At present, no imaging modality is capable of in vivo diagnosis of subepithelial airway wall pathology as signs of intubation-related injury. OBJECTIVES: To use Fourier domain long-range optical coherence tomography (LR-OCT) to acquire micrometer-resolution images of the airway wall of intubated neonates in a neonatal intensive care unit setting and to analyze images for histopathology and airway wall thickness. METHODS: LR-OCT of the neonatal laryngotracheal airway was performed a total of 94 times on 72 subjects (age, 1-175 d; total intubation, 1-104 d). LR-OCT images of the airway wall were analyzed in MATLAB. Medical records were reviewed retrospectively for extubation outcome. MEASUREMENTS AND MAIN RESULTS: Backward stepwise regression analysis demonstrated a statistically significant association between log(duration of intubation) and both laryngeal (P < 0.001; multiple r(2) = 0.44) and subglottic (P < 0.001; multiple r(2) = 0.55) airway wall thickness. Subjects with positive histopathology on LR-OCT images had a higher likelihood of extubation failure (odds ratio, 5.9; P = 0.007). Longer intubation time was found to be significantly associated with extubation failure. CONCLUSIONS: LR-OCT allows for high-resolution evaluation and measurement of the airway wall in intubated neonates. Our data demonstrate a positive correlation between laryngeal and subglottic wall thickness and duration of intubation, suggestive of progressive soft tissue injury. LR-OCT may ultimately aid in the early diagnosis of postintubation subglottic injury and help reduce the incidences of failed extubation caused by subglottic edema or acquired subglottic stenosis in neonates. Clinical trial registered with www.clinicaltrials.gov (NCT 00544427).
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Intubación Intratraqueal/efectos adversos , Laringoestenosis/diagnóstico , Tomografía de Coherencia Óptica/métodos , Diagnóstico Precoz , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Estudios RetrospectivosRESUMEN
Biofilm formation has been linked to ventilator-associated pneumonia, which is a prevalent infection in hospital intensive care units. Currently, there is no rapid diagnostic tool to assess the degree of biofilm formation or cellular biofilm composition. Optical coherence tomography (OCT) is a minimally invasive, nonionizing imaging modality that can be used to provide high-resolution cross-sectional images. Biofilm deposited in critical care patients' endotracheal tubes was analyzed in vitro. This study demonstrates that OCT could potentially be used as a diagnostic tool to analyze and assess the degree of biofilm formation and extent of airway obstruction caused by biofilm in endotracheal tubes.
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Biopelículas , Endoscopía/métodos , Contaminación de Equipos , Intubación Intratraqueal/efectos adversos , Tomografía de Coherencia Óptica/métodos , Artefactos , Cuidados Críticos , Infección Hospitalaria/diagnóstico , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Imagenología Tridimensional/métodos , Unidades de Cuidados Intensivos , Interferometría/métodos , Microscopía Electrónica de Rastreo , Fenotipo , Neumonía Asociada al Ventilador/diagnósticoRESUMEN
We report on the feasibility of using long-range swept-source optical coherence tomography (OCT) to detect airway changes following smoke inhalation in a sheep model. The long-range OCT system (with axial imaging range of 25 mm) and probe are capable of rapidly obtaining a series of high-resolution full cross-sectional images and three-dimensional reconstructions covering 20-cm length of tracheal and bronchial airways with airway diameter up to 25 mm, regardless of the position of the probe within the airway lumen. Measurements of airway thickness were performed at baseline and postinjury to show mucosal thickness changes following smoke inhalation.
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Imagenología Tridimensional/métodos , Lesión por Inhalación de Humo/diagnóstico , Tomografía de Coherencia Óptica/métodos , Animales , Bronquios/patología , Broncoscopía , Mucosa Respiratoria/patología , Ovinos , Lesión por Inhalación de Humo/patologíaRESUMEN
Many diseases involve changes in the biomechanical properties of tissue, and there is a close correlation between tissue elasticity and pathology. We report on the development of a phase-resolved acoustic radiation force optical coherence elastography method (ARF-OCE) to evaluate the elastic properties of tissue. This method utilizes chirped acoustic radiation force to produce excitation along the sample's axial direction, and it uses phase-resolved optical coherence tomography (OCT) to measure the vibration of the sample. Under 500-Hz square wave modulated ARF signal excitation, phase change maps of tissue mimicking phantoms are generated by the ARF-OCE method, and the resulting Young's modulus ratio is correlated with a standard compression test. The results verify that this technique could efficiently measure sample elastic properties accurately and quantitatively. Furthermore, a three-dimensional ARF-OCE image of the human atherosclerotic coronary artery is obtained. The result indicates that our dynamic phase-resolved ARF-OCE method can delineate tissues with different mechanical properties.
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Diagnóstico por Imagen de Elasticidad/métodos , Técnicas Fotoacústicas/métodos , Tomografía de Coherencia Óptica/métodos , Fenómenos Biomecánicos , Enfermedad de la Arteria Coronaria/patología , Enfermedad de la Arteria Coronaria/fisiopatología , Vasos Coronarios/patología , Vasos Coronarios/fisiopatología , Módulo de Elasticidad , Diagnóstico por Imagen de Elasticidad/instrumentación , Humanos , Fenómenos Ópticos , Fantasmas de Imagen , Técnicas Fotoacústicas/instrumentación , Tomografía de Coherencia Óptica/instrumentaciónRESUMEN
The traditional phase-resolved Doppler method demonstrates great success for in-vivo imaging of blood flow and blood vessels. However, the phase-resolved method always requires high phase stability of the system. In phase instable situations, the performance of the phase-resolved methods will be degraded. We propose a modified Doppler variance algorithm that is based on the intensity or amplitude value. Performances of the proposed algorithm are compared with traditional phase-resolved Doppler variance and color Doppler methods for both phase stable and phase instable systems. For the phase instable situation, the proposed algorithm demonstrates images without phase instability induced artifacts. In-vivo imaging of window-chamber hamster skin is demonstrated for phase instable situation with a spectrometer-based Fourier domain OCT system. A microelectromechanical systems (MEMS) based swept source OCT (SSOCT) system is also used to demonstrate the performance of the proposed method in a phase instable situation. The phase stability of the SSOCT system is analyzed. In-vivo imaging of the blood vessel of human skin is demonstrated with the proposed method and the SSOCT system. For the phase stable situation, the proposed algorithm also demonstrates comparable performance with traditional phase-resolved methods. In-vivo imaging of the human choroidal blood vessel network is demonstrated with the proposed method under the phase stable situation. Depth-resolved fine choroidal blood vessel networks are shown.
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Algoritmos , Efecto Doppler , Tomografía de Coherencia Óptica/métodos , Animales , Cricetinae , Análisis de Fourier , Humanos , Transición de Fase , Epitelio Pigmentado de la Retina/anatomía & histología , Análisis EspectralRESUMEN
Measuring the kinetic constants of protein-protein interactions at ultralow concentrations becomes critical in characterizing biospecific affinity, and exploring the feasibility of clinical diagnosis with respect to detection sensitivity, efficiency and accuracy. In this study, we propose a method that can calculate the binding constants of protein-protein interactions in sandwich assays at ultralow concentrations at the pg/mL level, using a localized surface plasmon coupled fluorescence fiber-optic biosensor (LSPCF-FOB). We discuss a two-compartment model to achieve reaction-limited kinetics under the stagnant conditions of the reaction chamber. The association rate constant, dissociation rate constant, and the equilibrium dissociation constant, that is, k(a), k(d), K(D), respectively, of the kinetics of binding between total prostate-specific antigen (t-PSA) and anti-t-PSA at concentrations from 0.1 pg/mL to 1 ng/mL, were measured either in PBS or in human serum. This is the first time that k(a), k(d), and K(D) have been measured at such a low concentration range in a complex sample such as human serum.
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Técnicas Biosensibles/métodos , Oro/química , Nanopartículas del Metal/química , Animales , Anticuerpos Inmovilizados/inmunología , Técnicas Biosensibles/instrumentación , Bovinos , Humanos , Inmunoglobulina G/inmunología , Cinética , Límite de Detección , Ratones , Fibras Ópticas , Unión Proteica , Espectrometría de Fluorescencia , Especificidad por SustratoRESUMEN
The spherical aberration induced by refractive-index mismatch results in the degradation on the quality of sectioning images in conventional confocal laser scanning microscope (CLSM). In this research, we have derived the theory of image formation in a Zeeman laser scanning confocal microscope (ZLSCM) and conducted experiments in order to verify the ability of reducing spherical aberration in ZLSCM. A Zeeman laser is used as the light source and produces the linearly polarized photon-pairs (LPPP) laser beam. With the features of common-path propagation of LPPP and optical heterodyne detection, ZLSCM shows the ability of reducing the specimen-induced spherical aberration and improving the axial resolution simultaneously.
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A dual-frequency equal-amplitude paired polarization heterodyne polarimeter (DEPHP) was set up in order to precisely measure the mutarotation rate constants of D-glucose in tridistilled water. The DEPHP is based on a balanced detector detection scheme for measurement of the optical rotation angle of D-glucose/water solution during the conversion process between alpha-D-glucose and beta-D-glucose while in a nonequilibrium state. The DEPHP can perform shot-noise-limited detection, so that the total optical rotation angle together with the mutarotation rate constants of alpha-D-glucose and beta-D-glucose conversion can be measured with high sensitivity. In this experiment, the sensitivity of the optical rotation angle measurement was 8.3 x 10(-5) deg/cm, while the total (k), forward (k(1)) and reverse (k(2)) mutarotation rate constants of D-glucose were found to be k = 7.67 x 10(-5) s(-1), k(1) = 2.76 x 10(-5) s(-1), and k(2) = 4.91 x 10(-5) s(-1), respectively, in tridistilled water. Moreover, using the DEPHP, we can measure the specific rotation angles of alpha-D-glucose and beta-D-glucose in water at 632.8 nm. They were 105 degrees and 12 degrees, respectively. Finally, the detection sensitivity of the DEPHP system is also discussed.
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Glucosa/química , Agua/química , Cinética , Rotación Óptica , Reproducibilidad de los ResultadosRESUMEN
This research proposed a dual-frequency heterodyne ellipsometer (DHE) in which a dual-frequency collinearly polarized laser beam with equal amplitude and zero phase difference between p- and s-polarizations is setup. It is based on the polarizer-sample-analyzer, PSA configuration of the conventional ellipsometer. DHE enables to characterize a generalized elliptical phase retarder by treating it as the combination of a linear phase retarder and a polarization rotator. The method for measuring elliptical birefringence of an elliptical phase retarder based on the equivalence theorem of an unitary optical system was derived and the experimental verification by use of DHE was demonstrated too. The experimental results show the capability of DHE on characterization of a generalized phase retardation plate accurately.