Corneal evaluation in ophthalmology necessitates cellular-resolution and fast imaging techniques allowing accurate diagnoses. Currently, the fastest volumetric imaging technique is Fourier-domain full-field optical coherence tomography (FD-FF-OCT) that uses a fast camera and a rapidly tunable laser source. Here, we demonstrate high-resolution, high-speed, non-contact corneal volumetric imaging in vivo with FD-FF-OCT that can acquire a single 3D volume with a voxel rate of 7.8 GHz. The spatial coherence of the laser source was suppressed to prevent it from focusing to a spot on the retina, and therefore, exceeding the maximum permissible exposure (MPE). Inherently volumetric nature of FD-FF-OCT data enabled flattening of curved corneal layers. Acquired FD-FF-OCT images revealed corneal cellular structures, such as epithelium, stroma and endothelium, as well as subbasal and mid-stromal nerves.
Purpose. To demonstrate accurate measurement of corneal elastic moduli in vivo with non-contact and non-invasive optical coherence elastography. Methods. Elastic properties (in-plane Young's modulus E and both in-plane, u, and out-of-plane, G, shear moduli) of rabbit cornea were quantified in vivo using non-contact dynamic Acoustic micro-Tapping Optical Coherence Elastography (AuT-OCE). The IOP-dependence of measured mechanical properties was explored in extracted whole globes following in vivo measurement. A nearly-incompressible transverse isotropic (NITI) model was used to reconstruct modul…
Purpose: To evaluate changes in the anisotropic elastic properties of ex vivo human cornea treated with UV cross-linking (CXL) using non-contact acoustic micro-tapping Optical Coherence Elastography (AuT-OCE) Design: AuT performed on normal and CXL ex vivo human donor cornea Methods: Elastic properties of normal and UV CXL treated human corneas were quantified using non-contact acoustic micro-tapping Optical Coherence Elastography (AuT-OCE) Main Outcome Measures: Corneal elastic moduli (in-plane Young's, E, and out-of-plane shear, G) can be evaluated in both normal and CXL treated tissues, as …
Mounting evidence connects the biomechanical properties of tissues to the development of eye diseases such as keratoconus, a common disease in which the cornea thins and bulges into a conical shape. However, measuring biomechanical changes in vivo with sufficient sensitivity for disease detection has proved challenging. Here, we present a first large-scale study (~200 subjects, including normal and keratoconus patients) using Brillouin light-scattering microscopy to measure longitudinal modulus in corneal tissues with high sensitivity and spatial resolution. Our results in vivo provide evidenc…
High-resolution optical imaging methods, such as confocal microscopy and full-field optical coherence tomography, capture flat optical sections of the sample. If the sample is curved, the optical field sections through several sample layers and the view of each layer is reduced. Here we present curved-field optical coherence tomography, capable of capturing optical sections of arbitrary curvature. We test the device on a challenging task of imaging the human cornea in vivo and achieve 10x larger viewing area comparing to the clinical state-of-the-art. This enables more precise cell and nerve c…
Possible depth-resolved reconstruction of shear moduli in the cornea following collagen crosslinking (CXL) with optical coherence tomography and elastography
Optical Coherence Tomography (OCT) is a non-invasive depth resolved optical imaging modality, that enables high resolution, cross-sectional imaging in biological tissues and materials at clinically relevant depths. Though OCT offers high resolution imaging, the best ultra-high-resolution OCT systems are limited to imaging structural changes with a resolution of one micron on a single B-scan within very limited depth. Nanosensitive OCT (nsOCT) is a recently developed technique that is capable of providing enhanced sensitivity of OCT to structural changes. Improving the sensitivity of OCT to det…
Possible depth-resolved reconstruction of shear moduli in the cornea following collagen crosslinking (CXL) with optical coherence tomography and elastography
提出结合 OCT 相位去相关与 AμT OCE 及双层模型，在离体人角膜中重建 CXL 后刚度的深度分布并量化手术效果。
Corneal cross-linking (CXL) using UVA irradiation with a riboflavin photosensitizer has grown from an interesting concept to a practical clinical treatment for corneal ectatic diseases globally, such as keratoconus. To characterize the corneal structural changes, existing methods such as X-ray microscopy, transmission electron microscopy (TEM), histology and optical coherence tomography have been used. However, these methods have various drawbacks such as invasive detection, the impossibility for in vivo measurement, or limited resolution and sensitivity to structural alterations. Here, we rep…
Glaucoma remains among the leading causes of blindness despite many treatment options available today. Effective treatment requires early diagnosis, which is difficult to achieve with existing imaging technologies that detect the already inflicted irreversible posterior eye damage. One improved approach could be to detect the changes in trabecular meshwork of the anterior eye that precede the intraocular pressure buildup and vision loss. Here we report a device that enables in vivo exploration of trabecular meshwork at cell resolution. The instrument combines two orthogonal views: en face view…
When the cornea becomes hydrated above its physiologic level it begins to significantly scatter light, loosing transparency and thus impairing eyesight. This condition, known as corneal edema, can be associated with different causes, such as corneal scarring, corneal infection, corneal inflammation, and others, making it difficult to diagnose and quantify. Previous works have shown that Second Harmonic Generation Microscopy (SHG) represents a valuable non-linear optical imaging tool to non-invasively identify and monitor changes in the collagen architecture of the cornea, potentially playing a…
Multiple light scattering in tissue limits the penetration of optical coherence tomography (OCT) imaging. Here, we present in-vivo OCT imaging of a live mouse using wavefront shaping to enhance the penetration depth. A digital micro-mirror device (DMD) was used in a spectral-domain OCT system for complex wavefront shaping of an incident beam which resulted in the optimal delivery of light energy into deep tissue. Ex-vivo imaging of chicken breasts and mouse ear tissues showed enhancements in the strength of the image signals and the penetration depth, and in-vivo imaging of the tail of a live …
We have developed a multimodal photoacoustic remote sensing (PARS) microscope combined with swept source optical coherence tomography for in vivo, non-contact retinal imaging. Building on the proven strength of multiwavelength PARS imaging, the system is applied for estimating retinal oxygen saturation in the rat retina. The capability of the technology is demonstrated by imaging both microanatomy and the microvasculature of the retina in vivo. To our knowledge this is the first time a non-contact photoacoustic imaging technique is employed for in vivo oxygen saturation measurement in the reti…
Anterior uveitis is the most common form of intraocular inflammation, and one of its main signs is the presence of white blood cells (WBCs) in the anterior chamber (AC). Clinically, the true composition of cells can currently only be obtained using AC paracentesis, an invasive procedure to obtain AC fluid requiring needle insertion into the AC. We previously developed a spectroscopic optical coherence tomography (SOCT) analysis method to differentiate between populations of RBCs and subtypes of WBCs, including granulocytes, lymphocytes and monocytes, both in vitro and in ACs of excised porcine…
In today's clinics, a cellular-resolution view of the cornea can be achieved only with an in vivo confocal microscope (IVCM) in contact with the eye. Here, we present a common-path Full-field/Spectral-domain OCT microscope (FF/SD OCT), which, for the first time, enables cell-detail imaging of the entire ocular surface (central and peripheral cornea, limbus, sclera, tear film) without contact and in real time. The device, which has been successfully tested in human subjects, is now ready for direct implementation in clinical research. Real-time performance is achieved through rapid axial eye tr…
Eindhoven University of Technology（埃因霍温理工大学）；Rigshospitalet - Glostrup（哥本哈根大学医院格洛斯楚普院区）；Copenhagen Eye Foundation（哥本哈根眼科基金会）
Corneal thickness (pachymetry) maps can be used to monitor restoration of corneal endothelial function, for example after Descemet's membrane endothelial keratoplasty (DMEK). Automated delineation of the corneal interfaces in anterior segment optical coherence tomography (AS-OCT) can be challenging for corneas that are irregularly shaped due to pathology, or as a consequence of surgery, leading to incorrect thickness measurements. In this research, deep learning is used to automatically delineate the corneal interfaces and measure corneal thickness with high accuracy in post-DMEK AS-OCT B-scan…
Copper phosphate micro-flowers coated with indocyanine green and iron oxide nanoparticles for in vivo localization optoacoustic tomography and magnetic actuation
Early diagnosis of ocular diseases improves the understanding of pathophysiology and helps with accurate monitoring and effective treatment. Advanced multimodal ocular imaging platforms play a crucial role in the visualization of the ocular components and provide clinicians with a valuable tool for evaluating different eye diseases. Here, for the first time, we present a non-contact, multimodal photoacoustic remote sensing (PARS) microscopy and swept-source optical coherence tomography (SS-OCT) for in-vivo functional and structural imaging of the eye. The system provides complementary imaging …
The penetration depth of photoacoustic imaging in biological tissues has been fundamentally limited by the strong optical attenuation when light is delivered externally through the tissue surface. To address this issue, we previously reported internal-illumination photoacoustic imaging using a customized radial-emission optical fiber diffuser, which, however, has complex fabrication, high cost, and non-uniform light emission. To overcome these shortcomings, we have developed a new type of low-cost fiber diffusers based on a graded-scattering method in which the optical scattering of the fiber …
Non-invasive functional imaging of molecular and cellular processes of vision is expected to have immense impact on research and clinical diagnostics. Although suitable intrinsic optical signals (IOS) have been observed ex vivo and in immobilized animals in vivo, it was so far not possible to obtain convincing IOS of photoreceptor activity in humans in vivo. Here, we observed spatially and temporally clearly resolved changes in the optical path length of the photoreceptor outer segment as response to an optical stimulus in living human. To obtain these changes, we evaluated phase data of a par…
Optical coherence tomography (OCT) is a powerful biomedical imaging technology that relies on the coherent detection of backscattered light to image tissue morphology in vivo. As a consequence, OCT is susceptible to coherent noise (speckle noise), which imposes significant limitations on its diagnostic capabilities. Here we show a method based purely on light manipulation that is able to entirely remove the speckle noise originating from turbid samples without any compromise in resolution. We refer to this method as Speckle-Free OCT (SFOCT). Using SFOCT, we succeeded in revealing small structu…
Research and medicine rely on non-invasive optical techniques to image living tissue with high resolution in space and time. But so far a single data acquisition could not provide entirely diffraction-limited tomographic volumes of rapidly moving or changing targets, which additionally becomes increasingly difficult in the presence of aberrations, e.g., when imaging retina in vivo. 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.…
We demonstrate in vivo endoscopic optical coherence tomography (OCT) imaging in the forward direction using a flexible fiber bundle. In comparison to current conventional forward looking probe schemes, our approach simplifies the endoscope design by avoiding the integration of any beam steering components in the distal probe end due to 2D scanning of a focused light beam over the proximal fiber bundle surface. We describe the challenges that arise when OCT imaging with a fiber bundle is performed, such as multimoding or cross-coupling. The performance of different fiber bundles with varying pa…
Imaging airway tissue, optical coherence tomography (OCT) provides cross-sectional images of tissue structures, shows cilia movement and mucus secretion, but does not provide sufficient contrast to differentiate individual cells. By using fast sequences of microscopic resolution OCT (mOCT) images, OCT can picture small signal fluctuations (dynamic) OCT and overcomes the lack in contrast caused by speckle noise. In this way, OCT visualizes airway morphology on a cellular level and allows to track the dynamic behavior of immune cells, as well as mucus transport and secretion. Here, we demonstrat…
Purpose: To assess the early-phase of tear clearance rate (TCR) with anterior segment optical coherence tomography (OCT) and to determine the association between TCR and other clinical measures of the tear film in a group of young subjects with different levels of tear film quality. Methods: TCR was classified as the percentage decrease of the inferior tear meniscus height 30s after instillation of 5uL 0.9% saline solution. Fifty subjects (32F and 18M) aged (mean +/- standard deviation) 25.5 +/- 4.3 years volunteered for the study. It consisted of a review of medical history, Ocular Surface Di…
Although cone-beam CT (CBCT) has been used to guide irradiation for pre-clinical radiotherapy(RT) research, it is limited to localize soft tissue target especially in a low imaging contrast environment. Knowledge of target shape is a fundamental need for RT. Without such information to guide radiation, normal tissue can be irradiated unnecessarily, leading to experimental uncertainties. Recognition of this need led us to develop quantitative bioluminescence tomography (QBLT), which provides strong imaging contrast to localize optical targets. We demonstrated its capability of guiding conformal…
Eye movements are commonly seen as an obstacle to high-resolution ophthalmic imaging. In this context we study the natural axial movements of the in vivo human eye and show that they can be used to modulate the optical phase and retrieve tomographic images via time-domain full-field optical coherence tomography (TD-FF-OCT). This approach opens a path to a simplified ophthalmic TD-FF-OCT device, operating without the usual piezo motor-camera synchronization. The device demonstrates in vivo human corneal images under the different image retrieval schemes (2-phase and 4-phase) and different expos…
A polarization-multiplexed, dual-beam setup is proposed to expand the field of view for a swept source optical coherence tomography angiography (OCTA) system. This method used a Wollaston prism to split sample path light into two orthogonal-polarized beams. This allowed two beams to shine on the cornea at an angle separation of ~ 14 degrees, which led to a separation of ~ 4.2 mm on the retina. A 3-mm glass plate was inserted into one of the beam paths to set a constant path length difference between the two polarized beams so the interferogram from the two beams are coded at different frequenc…
Recent studies have shown potential for using polarisation sensitive optical coherence tomography (PS-OCT) to study cartilage morphology, and to be potentially used as an in-vivo, non-invasive tool for detecting osteoarthritic changes. However, there has been relatively limited ability of this method to quantify the subtle changes that occur in the early stages of cartilage degeneration. An established mechanical indenting technique that has previously been used to examine the microstructural response of articular cartilage was employed to fix the bovine samples in an indented state. The sampl…
We developed a new endogenous approach to reveal subcellular metabolic contrast in fresh ex vivo tissues taking advantage of the time dependence of the full field optical coherence tomography interferometric signals. This method reveals signals linked with local activity of the endogenous scattering elements which can reveal cells where other imaging techniques fail or need exogenous contrast agents. We benefit from the micrometric transverse resolution of full field OCT to image intracellular features. We used this time dependence to identify different dynamics at the millisecond scale on a w…
Angiogenesis, the formation of new vessels, is one of the key mechanisms in tumor development and an appealing target for therapy. Non-invasive, high-resolution, high sensitivity, quantitative 3D imaging techniques are required to correctly depict tumor heterogeneous vasculature over time. Ultrafast Doppler was recently introduced and provides an unprecedented combination of resolution, penetration depth and sensitivity without requiring any contrast agents. The technique was further extended to 3D with Ultrafast Doppler Tomography (UFD-T). In this work, UFD-T was applied to the monitoring of …
Optical coherence tomography (OCT) angiography is a well-established in vivo imaging technique to assess the overall vascular morphology of tissues and is an emerging field of research for the assessment of blood flow dynamics and functional parameters such as oxygen saturation. In this study, we present a modified scanning based correlation mapping OCT (cmOCT) using a 200 kHz high speed swept source OCT system operating at 1300 nm and demonstrate its wide field imaging capability in ocular angiographic studies.
The in vivo observation of the human retina at the cellular level is crucial to detect lesions before irreversible visual loss occurs, to follow the time course of retinal diseases and to evaluate and monitor the early effects of treatments. Despite the phenomenal advances in optical coherence tomography (OCT) and adaptive optics systems, in vivo imaging of several retinal cells is still elusive. Here we propose a radically different method compared to OCT, called transscleral optical phase imaging (TOPI), which allows to image retinal cells with high contrast, high resolution, and within an a…
Forward-viewing endoscopic optical coherence tomography (OCT) provides 3D imaging in vivo, and can be combined with widefield fluorescence imaging by use of a double-clad fiber. However, it is technically challenging to build a high-performance miniaturized 2D scanning system with a large field-of-view. In this paper we demonstrate how a 1D scanning probe, which produces cross-sectional OCT images (B-scans) and 1D fluorescence T-scans, can be transformed into a 2D scanning probe by manual scanning along the second axis. OCT volumes are assembled from the B-scans using speckle decorrelation mea…
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 sig…
We develop a new three-dimensional time-dependent radiative transfer code, TRINITY (Time-dependent Radiative transfer In Near-Infrared TomographY), for in-vivo diffuse optical tomography (DOT). The simulation code is based on the design of long radiation rays connecting boundaries of a computational domain, which allows us to calculate light propagation with little numerical diffusion. We parallelize the code with Message Passing Interface (MPI) using the domain decomposition technique and confirm the high parallelization efficiency, so that simulations with a spatial resolution of $\sim 1$ mi…
One of the greatest needs in modern ophthalmology is access to the eye choroid in vivo. Without it, it is difficult to introduce new therapies and to understand most of the pathological changes in the eye, because of the total blood flow in the eye is caused by the choroidal circulation. It is entirely responsible for the metabolism of photoreceptors, which each of us feels every day and senses when its deficits begin. Optical selection of choroid and choriocapillaris by currently used imaging techniques such as indocyanine angiography, scanning laser ophthalmoscopy or optical coherence tomogr…
In vivo real-time epifluorescence imaging of mouse hindlimb vasculatures in the second near-infrared region (NIR-II, 1.1~1.4 microns) is performed using single-walled carbon nanotubes (SWNTs) as fluorophores. Both high spatial resolution (~30 microns) and temporal resolution (<200 ms/frame) for small vessel imaging are achieved 1-3 mm deep in the tissue owing to the beneficial NIR-II optical window that affords deep anatomical penetration and low scattering. This spatial resolution is unattainable by traditional NIR imaging (NIR-I, 0.75~0.9 microns) or microscopic computed tomography (micro-CT…
This paper reports on advances in optical coherence tomography (OCT) for application in dermatology. Full-field OCT is a particular approach of OCT based on white-light interference microscopy. FF-OCT produces en face tomographic images by arithmetic combination of interferometric images acquired with an area camera and by illuminating the whole field of view with low-coherence light. The major interest of FF-OCT lies in its high imaging spatial resolution ($\sim$ 1.0 $μ$m) in both lateral and axial directions, using a simple and robust experimental arrangement. Line-field OCT (LF-OCT) is a re…
Dynamic optical coherence elastography (OCE) tracks mechanical wave propagation in the subsurface region of tissue to image its shear modulus. For bulk shear waves, the lateral resolution of the reconstructed modulus map (i.e., elastographic resolution) can approach optical coherence tomography (OCT) capabilities, typically a few tens of microns. Here we perform comprehensive numerical simulations and acoustic micro-tapping OCE experiments to show that for the typical situation of guided wave propagation in bounded media, such as cornea, the elastographic resolution cannot reach the OCT resolu…
There is an increasing need for label free methods that could reveal intracellular structures and dynamics. In this context, we develop a new optical tomography method working in transmission - Full-field optical transmission tomography (FF-OTT). The method can measure the forward scattering signals and reveal the metabolic time-dependent signals in living cells. FF-OTT is a common path interferometer taking advantage of the Gouy phase shift - a π phase shift that the light wave experiences around the focus. By modulating position of the focus one can alter the phase of the scattered light. De…
There has been a longstanding expectation that the optical resolution embodiment of photoacoustic tomography could have a substantial impact on gastrointestinal endoscopy by enabling microscopic visualization of the vasculature based on the endogenous contrast mechanism. Although multiple studies have demonstrated the in vivo imaging capability of a developed imaging device over the last decade, the implementation of such an endoscopic system that can be applied immediately when necessary via the instrument channel of a video endoscope has been a challenge. In this study, we developed a 3.38 m…
The health of cells found in the inner retinal layers of the human eye is crucial to understand the onset of diseases of the retina such as macular degeneration and retinopathy. A challenge is to periodically image these cells in human eyes to detect abnormalities well before physiological and pathological changes occur. However, in vivo imaging of many of these cells is still elusive despite the phenomenal advances in Optical Coherence Tomography (OCT) and Adaptive Optics systems. It stems from the fact that cell contrast in reflection is extremely low. Here, we report a major advance towards…
