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Holographic optical coherence tomography (OCT) is a powerful imaging technique, but its ability to reveal low-reflectivity features is limited. In this study, we performed holographic OCT by incoherently averaging volumes with changing diffuse illumination of numerical aperture (NA) equal to the detection NA. While the reduction of speckle from singly scattered light is only modest, we discovered that speckle from multiply scattered light can be arbitrarily reduced, resulting in substantial improvements in image quality. This technique also offers the advantage of suppressing noises arising from spatial coherence, and can be implemented with a partially spatially incoherent light source for further mitigation of multiple scattering. Finally, we show that although holographic reconstruction capabilities are increasingly lost with decreasing spatial coherence, they can be retained over an axial range sufficient to standard OCT applications.
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Full-field swept-source optical coherence tomography (FF-SS-OCT) and laser Doppler holography (LDH) are two holographic imaging techniques presenting unique capabilities for ophthalmology. We report on interlaced FF-SS-OCT and LDH imaging with a single instrument. Effectively, retinal blood flow and pulsation could be quasi-simultaneously monitored. This instrument holds potential for a wide scope of ophthalmic applications.
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Holografia , Tomografia de Coerência Óptica , Angiografia , Lasers , Retina/diagnóstico por imagem , Tomografia de Coerência Óptica/métodosRESUMO
In optical coherence tomography (OCT), lateral motion is determined either by speckle tracking or by multi-aperture Doppler OCT. Here we show that both methods may provide incorrect results because, outside the focal plane, non-uniform axial motion is misinterpreted as lateral motion. First, we demonstrate the existence of this artifact by means of a simulation for speckle tracking. Then the physical origin of the artifact and its mathematical relation to defocus and axial motion are explained. It is shown that speckle tracking and multi-aperture Doppler OCT are equally affected by the artifact, which has a considerable effect, even for a defocus of less than one Rayleigh length.
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Computational adaptive optics (CAO) is emerging as a viable alternative to hardware-based adaptive optics-in particular when applied to optical coherence tomography of the retina. For this technique, algorithms are required that detect wavefront errors precisely and quickly. Here we propose an extension of the frequently used subaperture image correlation. By applying this algorithm iteratively and, more importantly, comparing each subaperture not to the central subaperture but to several randomly selected apertures, we improved aberration correction. Since these modifications only slightly increase the run time of the correction, we believe this method can become the algorithm of choice for many CAO applications.
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Functional retinal imaging, especially of neuronal activity non-invasively in humans, is of tremendous interest. Although the activation of photoreceptor cells (PRCs) could be detected in humans, imaging the function of other retinal neurons had been so far hardly possible. Here, using phase-sensitive full-field swept-source optical coherence tomography (FF-SS-OCT), we show simultaneous imaging of the activation in the photoreceptor and ganglion cell layer/inner plexiform layer (GCL/IPL). The signals from the GCL/IPL are 10-fold smaller than those from the PRC and were detectable only using algorithms for suppression of motion artifacts and pulsatile blood flow in the retinal vessels. FF-SS-OCT with improved phase evaluation algorithms, therefore, allowed us to map functional connections between PRC and GCL/IPL, confirming previous ex vivo results. The demonstrated functional imaging of retinal neuronal layers can be a valuable tool in diagnostics and basic research.
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Neurônios/citologia , Células Fotorreceptoras de Vertebrados/citologia , Tomografia de Coerência Óptica , Sobrevivência Celular , Humanos , Fatores de TempoRESUMO
Noninvasive functional imaging of molecular and cellular processes of vision may have immense impact on research and clinical diagnostics. Although suitable intrinsic optical signals (IOSs) have been observed ex vivo and in immobilized animals in vivo, detecting IOSs of photoreceptor activity in living humans was cumbersome and time consuming. Here, we observed clear spatially and temporally resolved changes in the optical path length of the photoreceptor outer segment as a response to an optical stimulus in the living human eye. To witness these changes, we evaluated phase data obtained with a parallelized and computationally aberration-corrected optical coherence tomography system. The noninvasive detection of optical path length changes shows neuronal photoreceptor activity of single cones in living human retina, and therefore, it may provide diagnostic options in ophthalmology and neurology and could provide insights into visual phototransduction in humans.
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Retina/diagnóstico por imagem , Retina/fisiologia , Células Fotorreceptoras Retinianas Cones/fisiologia , Humanos , Fenômenos Ópticos , Estimulação Luminosa , Tomografia de Coerência Óptica , Adulto JovemRESUMO
Doppler optical coherence tomography (OCT) quantifies axial motion with high precision, whereas lateral motion cannot be detected by a mere evaluation of phase changes. This problem was solved by the introduction of three-beam Doppler OCT, which, however, entails a high experimental effort. Here, we present the numerical analogue to this experimental approach. Phase-stable complex-valued OCT datasets, recorded with full-field swept-source OCT, are filtered in the Fourier domain to limit imaging to different computational subapertures. These are used to calculate all three components of the motion vector with interferometric precision. As known from conventional Doppler OCT for axial motion only, the achievable accuracy exceeds the actual imaging resolution by orders of magnitude in all three dimensions. The feasibility of this method is first demonstrated by quantifying micro-rotation of a scattering sample. Subsequently, a potential application is explored by recording the 3D motion vector field of tissue during laser photocoagulation in ex-vivo porcine retina.
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Aberration-corrected imaging of human photoreceptor cells, whether hardware or software based, presently requires a complex and expensive setup. Here we use a simple and inexpensive off-axis full-field time-domain optical coherence tomography (OCT) approach to acquire volumetric data of an in vivo human retina. Full volumetric data are recorded in 1.3 s. After computationally correcting for aberrations, single photoreceptor cells were visualized. In addition, the numerical correction of ametropia is demonstrated. Our implementation of full-field optical coherence tomography combines a low technical complexity with the possibility for computational image correction.
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Processamento de Imagem Assistida por Computador , Retina/diagnóstico por imagem , Tomografia de Coerência Óptica/métodos , Algoritmos , Custos e Análise de Custo , Humanos , Fatores de Tempo , Tomografia de Coerência Óptica/economiaRESUMO
In numerous applications, Fourier-domain optical coherence tomography (FD-OCT) suffers from a limited imaging depth due to signal roll-off, a limited focal range, and autocorrelation noise. Here, we propose a parallel full-field FD-OCT imaging method that uses a swept laser source and an area camera in combination with an off-axis reference, which is incident on the camera at a small angle. As in digital off-axis holography, this angle separates autocorrelation signals and the complex conjugated mirror image from the actual signal in Fourier space. We demonstrate that by reconstructing the signal term only, this approach enables full-range imaging, i.e., it increases the imaging depth by a factor of two, and removes autocorrelation artifacts. The previously demonstrated techniques of inverse scattering and holoscopy can then numerically extend the focal range without loss of lateral resolution or imaging sensitivity. The resulting, significantly enhanced measurement depth is demonstrated by imaging a porcine eye over its entire depth, including cornea, lens, and retina. Finally, the feasibility of in vivo measurements is demonstrated by imaging the living human retina.
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With a simple setup, mainly composed of a low coherence light source and a camera, full-field optical coherence tomography (FF-OCT) allows volumetric tissue imaging. However, fringe washout constrains its use in retinal imaging. Here, we present a novel motion-insensitive approach to FF-OCT, which introduces path-length differences between the reference and the sample light in neighboring pixels using an off-axis reference beam. The temporal carrier frequency in scanned time-domain OCT is replaced by a spatial carrier frequency. Volumetric in-vivo FF-OCT measurements of the human retina were acquired in only 1.3 s, comparable to the acquisition times of current clinically used OCT devices.
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Retina/diagnóstico por imagem , Tomografia de Coerência Óptica/métodos , HumanosRESUMO
We demonstrate a new noninvasive method to assess biomechanical properties of the retinal vascular system. Phase-sensitive full-field swept-source optical coherence tomography (PhS-FF-SS-OCT) is used to investigate retinal vascular dynamics at unprecedented temporal resolution. The motion of retinal tissue that is induced by expansion of the vessels therein is measured with an accuracy of about 10 nm. The pulse shapes of arterial and venous pulsations, their temporal delays, as well as the frequency-dependent pulse propagation through the capillary bed, are determined. For the first time, imaging speed and motion sensitivity are sufficient for a direct measurement of pulse waves propagating with more than 600 mm/s in retinal vessels of a healthy young subject.
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Artéria Retiniana/fisiologia , Veia Retiniana/fisiologia , Tomografia de Coerência Óptica/métodos , Humanos , MovimentoRESUMO
In recent years, optoretinography has become an important functional imaging method for the retina, as light-evoked changes in the photoreceptors have been demonstrated for a large number of different OCT systems. Full-field swept-source optical coherence tomography (FF-SS-OCT) is particularly phase-stable, and it is currently the only technique sensitive enough to detect the smaller functional changes in the inner plexiform layer (IPL). However, the resolution of state-of-the art FF-SS-OCT systems is not high enough to distinguish individual photoreceptors. This makes it difficult to separate rods from cones. In this work, we circumvent this problem by separating the functional changes in rods and cones by their different temporal dynamics to the same light stimulus. For this purpose, a mathematical model was developed that represents the measured signals as a superposition of two impulse responses. The developed model describes the measured data under different imaging conditions very well and is able to analyze the sensitivity and temporal dynamics of the two photoreceptor types separately.
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Purpose: The lateral resolution of an optical coherence tomography (OCT) instrument was considered to be equal to the illumination spot size on the retina. To evaluate the potential lateral resolution of the Spectralis OCT, an instrument calculated to have a 14 µm resolution. Methods: The lateral point spread function (PSF) was evaluated using diamond abrasive powder 0 to 1 µm in diameter in silicone elastomer and a validated target with 800 nm FeO particles in urethane. The amplitude transfer function was calculated from human OCT images. Finally, resolution was measured using the 1951 USAF target. Results: Measurement of the lateral PSF from 1215 diamond particle images yielded a full-width half maximum (FWHM) to be 5.11 µm and for 732 FeO particles, 4.9 µm. From the amplitude transfer function, the FWHM of the diffraction limited PSF was calculated to be 5.0 µm. The USAF target imaging showed a lateral resolution of 4.6 µm. Conclusions: Although a calculation of the spot size of the illumination beam was reported in the past as the lateral resolution of the OCT instrument, the actual lateral resolution is better by a factor of at least 2.5 times. The clinically used A-scan spacing was derived from the calculated, and not the true resolution, and results in under sampling. This set of findings likely apply to all commercial clinical instruments. Translational Relevance: The scan density parameters of past and present commercial OCT instruments were based on earlier translational concepts, which now appear to have been incorrect.
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Retina , Tomografia de Coerência Óptica , HumanosRESUMO
Non-invasive spatially resolved functional imaging in the human retina has recently attracted considerable attention. Particularly functional imaging of bipolar and ganglion cells could aid in studying neuronal activity in humans, including an investigation of processes of the central nervous system. Recently, we imaged the activity of the inner neuronal layers by measuring nanometer-size changes of the cells within the inner plexiform layer (IPL) using phase-sensitive optical coherence tomography (OCT). In the IPL, there are connections between the neuronal cells that are dedicated to the processing of different aspects of the visual information, such as edges in the image or temporal changes. Still, so far, it was not possible to assign functional changes to single cells or cell classes in living humans, which is essential for studying the vision process. One characteristic of signal processing in the IPL is that different aspects of the visual impression are only processed in specific sub-layers (strata). Here, we present an investigation of these functional signals for three different sub-layers in the IPL with the aim to separate different properties of the visual signal processing. Whereas the inner depth-layer, closest to the ganglion cells, exhibits an increase in the optical path length, the outer depth-layer, closest to the bipolar cell layer, exhibits a decrease in the optical path length. Additionally, we found that the central depth is sensitive to temporal changes, showing a maximum response at a stimulation frequency of around 12.5 Hz. The results demonstrate that the signals from different cell types can be distinguished by phase-sensitive OCT.
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A method for numerical estimation and correction of aberrations of the eye in fundus imaging with optical coherence tomography (OCT) is presented. Aberrations are determined statistically by using the estimate based on likelihood function maximization. The method can be considered as an extension of the phase gradient autofocusing algorithm in synthetic aperture radar imaging to 2D optical aberration correction. The efficacy of the proposed method has been demonstrated in OCT fundus imaging with 6λ aberrations. After correction, single photoreceptors were resolved. It is also shown that wave front distortions with high spatial frequencies can be determined and corrected.
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Algoritmos , Tomografia de Coerência Óptica , Fundo de Olho , Funções VerossimilhançaRESUMO
Phase-sensitive coherent imaging exploits changes in the phases of backscattered light to observe tiny alterations of scattering structures or variations of the refractive index. But moving scatterers or a fluctuating refractive index decorrelate the phases and speckle patterns in the images. It is generally believed that once the speckle pattern has changed, the phases are scrambled and any meaningful phase difference to the original pattern is removed. As a consequence, diffusion and tissue motion that cannot be resolved, prevent phase-sensitive imaging of biological specimens. Here, we show that a phase comparison between decorrelated speckle patterns is still possible by utilizing a series of images acquired during decorrelation. The resulting evaluation scheme is mathematically equivalent to methods for astronomic imaging through the turbulent sky by speckle interferometry. We thus adopt the idea of speckle interferometry to phase-sensitive imaging in biological tissues and demonstrate its efficacy for simulated data and imaging of photoreceptor activity with phase-sensitive optical coherence tomography. We believe the described methods can be applied to many imaging modalities that use phase values for interferometry.
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This paper comments on the article "Retinal pulse wave velocity measurement using spectral-domain optical coherence tomography" by Qian Li et al. The authors propose a method to determine the pulse wave velocity in retinal arteries and veins. This method should enable a noninvasive determination of biomechanical properties of the vessel network, particularly the elasticity of the vessel walls. Although the observations the authors made might seem reasonable at first glance, they are in fact highly surprising and contradictory to theoretical predictions and previously published results.
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Artéria Retiniana , Tomografia de Coerência Óptica , Análise de Onda de Pulso , Retina , VeiasRESUMO
Full-field swept-source optical coherence tomography (FF-SS-OCT) was recently shown to allow new and exciting applications for imaging the human eye that were previously not possible using current scanning OCT systems. However, especially when using cameras that do not acquire data with hundreds of kHz frame rate, uncorrected phase errors due to axial motion of the eye lead to a drastic loss in image quality of the reconstructed volumes. Here we first give a short overview of recent advances in techniques and applications of parallelized OCT and finally present an iterative and statistical algorithm that estimates and corrects motion-induced phase errors in the FF-SS-OCT data. The presented algorithm is in many aspects adopted from the phase gradient autofocus (PGA) method, which is frequently used in synthetic aperture radar (SAR). Following this approach, the available phase errors can be estimated based on the image information that remains in the data, and no parametrization with few degrees of freedom is required. Consequently, the algorithm is capable of compensating even strong motion artifacts. Efficacy of the algorithm was tested on simulated data with motion containing varying frequency components. We show that even in strongly blurred data, the actual image information remains intact, and the algorithm can identify the phase error and correct it. Furthermore, we use the algorithm to compensate real phase error in FF-SS-OCT imaging of the human retina. Acquisition rates can be reduced by a factor of three (from 60 to 20 kHz frame rate) with an image quality that is even higher compared to uncorrected volumes recorded at the maximum acquisition rate. The presented algorithm for axial motion correction decreases the high requirements on the camera frame rate and thus brings FF-SS-OCT closer to clinical applications.
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Certain topics in research and advancements in medical diagnostics may benefit from improved temporal and spatial resolution during non-invasive optical imaging of living tissue. However, so far no imaging technique can generate entirely diffraction-limited tomographic volumes with a single data acquisition, if the target moves or changes rapidly, such as the human retina. Additionally, the presence of aberrations may represent further difficulties. We show that a simple interferometric setup-based on parallelized optical coherence tomography-acquires volumetric data with 10 billion voxels per second, exceeding previous imaging speeds by an order of magnitude. This allows us to computationally obtain and correct defocus and aberrations resulting in entirely diffraction-limited volumes. As demonstration, we imaged living human retina with clearly visible nerve fiber layer, small capillary networks, and photoreceptor cells. Furthermore, the technique can also obtain phase-sensitive volumes of other scattering structures at unprecedented acquisition speeds.
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Processamento de Imagem Assistida por Computador/estatística & dados numéricos , Imageamento Tridimensional/instrumentação , Interferometria/métodos , Imagem Óptica/instrumentação , Retina/ultraestrutura , Tomografia de Coerência Óptica/métodos , Capilares/anatomia & histologia , Capilares/ultraestrutura , Voluntários Saudáveis , Humanos , Imageamento Tridimensional/métodos , Interferometria/instrumentação , Fibras Nervosas/ultraestrutura , Imagem Óptica/métodos , Retina/anatomia & histologia , Fatores de Tempo , Tomografia de Coerência Óptica/instrumentaçãoRESUMO
A full-field speckle interferometry method for non-contact and prospectively high speed Photoacoustic Tomography is introduced and evaluated as proof of concept. Thermoelastic pressure induced changes of the objects topography are acquired in a repetitive mode without any physical contact to the object. In order to obtain high acquisition speed, the object surface is illuminated by laser pulses and imaged onto a high speed camera chip. In a repetitive triple pulse mode, surface displacements can be acquired with nanometre sensitivity and an adjustable sampling rate of e.g. 20 MHz with a total acquisition time far below one second using kHz repetition rate lasers. Due to recurring interferometric referencing, the method is insensitive to thermal drift of the object due to previous pulses or other motion. The size of the investigated area and the spatial and temporal resolution of the detection are scalable. In this study, the approach is validated by measuring a silicone phantom and a porcine skin phantom with embedded silicone absorbers. The reconstruction of the absorbers is presented in 2D and 3D. The sensitivity of the measurement with respect to the photoacoustic detection is discussed. Potentially, Photoacoustic Imaging can be brought a step closer towards non-anaesthetized in vivo imaging and new medical applications not allowing acoustic contact, such as neurosurgical monitoring or burnt skin investigation.