Gabor-domain Optical Coherence Microscopy Research Articles

Model-based optical coherence tomography angiography enables motion-insensitive vascular imaging.

We present a significant step toward ultrahigh-resolution, motion-insensitive characterization of vascular dynamics. Optical coherence tomography angiography (OCTA) is an invaluable diagnostic technology for non-invasive, label-free vascular imaging in vivo. However, since it relies on detecting moving cells from consecutive scans, high-resolution OCTA is susceptible to tissue motion, which imposes challenges in resolving and quantifying small vessels. We developed a novel OCTA technique named ultrahigh-resolution factor angiography (URFA) by modeling repeated scans as generative latent variables, with a common variance representing shared features and a unique variance representing motion. By iteratively maximizing the combined log-likelihood probability of these variances, the unique variance is largely separated. Meanwhile, features in the common variance are decoupled, in which vessels with dynamic flow are extracted from tissue structure by integrating high-order factors. Combined with Gabor-domain optical coherence microscopy, URFA successfully extracted high-resolution cutaneous vasculature despite severe involuntary tissue motion and scanner oscillation, significantly improving the visualization and characterization of micro-capillaries in vivo. Compared with the conventional approach, URFA reduces motion artifacts by nearly 50% on average, evaluated on local differences.

Spectral fusing Gabor domain optical coherence microscopy based on FPGA processing

High-resolution imaging using high numerical aperture imaging optics is commonly known to cause a narrow depth of focus, which limits the depth of field in optical coherence tomography (OCT). To achieve semi-invariant high resolution in all directions, Gabor domain optical coherence microscopy (GD-OCM) combines the in-focus regions of multiple cross-sectional images that are acquired while shifting the focal plane of the objective lens. As a result, GD-OCM requires additional processes for in-focus extraction and fusion, leading to longer processing times, as compared with conventional frequency domain OCT (FD-OCT). We previously proposed a method of spectral domain Gabor fusion that has been proven to improve the processing speed of GD-OCM. To investigate the full potential of the spectral domain Gabor fusion technique, we present the implementation of the spectral domain Gabor fusion algorithm using field programmable gate arrays (FPGAs) in a spectral acquisition hardware device. All filtering processes are now performed in an acquisition device as opposed to the post-processing of the original GD-OCM, which reduces the amount of data transfer between the image acquisition device and the processing host. To clearly demonstrate the imaging performance of the implemented system, we performed GD-OCM imaging of a stack of polymeric tapes. GD-OCM imaging was performed over seven focus zones. The results showed that the processing time for linear wavenumber calibration and spectral Gabor filtering can be improved with FPGA implementation. The total processing time was improved by about 35%.

Near-Histologic Resolution Images of Cervical Dysplasia Obtained With Gabor Domain Optical Coherence Microscopy.

ObjectiveHistopathology is the criterion standard for evaluating cervical squamous intraepithelial neoplasia (dysplasia). In this pilot feasibility study, we examined whether a novel 3-dimensional imaging device using Gabor-domain optical coherence microscopy (GDOCM) could distinguish features of cervical dysplasia comparable with histopathology.MethodsA prospective observational pilot study enrolled a small sample of women undergoing loop electrosurgical excision procedure for cervical squamous intraepithelial neoplasia. Fresh ex vivo specimens were imaged with the GDOCM device. Digital images were reviewed by a pathologist who was blinded to the histopathology results. Histopathologic features were then compared with the digital observations.ResultsStandard histologic features of cervical squamous epithelium and of squamous intraepithelial neoplasia could be observed in GDOCM images. Cervical epithelium, stroma, basement membrane, and squamous papilla could all be identified. Human papillomavirus effects, such as vacuolization and cellular density, were also observed.ConclusionsA GDOCM imaging system has the potential to obtain histologic resolution images of the cervix in the evaluation of squamous intraepithelial neoplasia. This pilot study allowed for optimizing the imaging system and paved the way for a future diagnostic accuracy study. The development of this technology could streamline the evaluation of patients at risk for cervical neoplasia.

Unbiased corneal tissue analysis using Gabor-domain optical coherence microscopy and machine learning for automatic segmentation of corneal endothelial cells.

.Significance: An accurate, automated, and unbiased cell counting procedure is needed for tissue selection for corneal transplantation.Aim: To improve accuracy and reduce bias in endothelial cell density (ECD) quantification by combining Gabor-domain optical coherence microscopy (GDOCM) for three-dimensional, wide field-of-view () corneal imaging and machine learning for automatic delineation of endothelial cell boundaries.Approach: Human corneas stored in viewing chambers were imaged over a wide field-of-view with GDOCM without contacting the specimens. Numerical methods were applied to compensate for the natural curvature of the cornea and produce an image of the flattened endothelium. A convolutional neural network (CNN) was trained to automatically delineate the cell boundaries using 180 manually annotated images from six corneas. Ten additional corneas were imaged with GDOCM and compared with specular microscopy (SM) to determine performance of the combined GDOCM and CNN to achieve automated endothelial counts relative to current procedural standards.Results: Cells could be imaged over a larger area with GDOCM than SM, and more cells could be delineated via automatic cell segmentation than via manual methods. ECD obtained from automatic cell segmentation of GDOCM images yielded a correlation of 0.94 () with the manual segmentation on the same images, and correlation of 0.91 () with the corresponding manually counted SM results.Conclusions: Automated endothelial cell counting on GDOCM images with large field of view eliminates selection bias and reduces sampling error, which both affect the gold standard of manual counting on SM images.

In vivo imaging of corneal nerves and cellular structures in mice with Gabor-domain optical coherence microscopy.

Gabor-domain optical coherence microscopy (GDOCM) demonstrated in vivo corneal imaging with cellular resolution and differentiation in mice over a field of view of 1 mm2. Contact and non-contact imaging was conducted on six healthy and six hyperglycemic C57BL/6J mice. Cellular resolution in the 3D GDOCM images was achieved after motion correction. Corneal nerve fibers were traced and their lengths and branches calculated. Noncontact, label-free imaging of corneal nerves has clinical utility in health and disease, and in transplant evaluation. To the authors' knowledge, this is the first report of in vivo 3D corneal imaging in mice with the capability to resolve nerve fibers using a non-contact imaging modality.

Gabor domain optical coherence microscopy combined with laser scanning confocal fluorescence microscopy.

We report on the development of fluorescence Gabor domain optical coherence microscopy (Fluo GD-OCM), a combination of GD-OCM with laser scanning confocal fluorescence microscopy (LSCFM) for synchronous micro-structural and fluorescence imaging. The dynamic focusing capability of GD-OCM provided the adaptive illumination environment for both modalities without any mechanical movement. Using Fluo GD-OCM, we imaged ex vivo DsRed-expressing cells in the brain of a transgenic mouse, as well as Cy3-labeled ganglion cells and Cy3-labeled astrocytes from a mouse retina. The self-registration of images taken by the two different imaging modalities showed the potential for a correlative study of subjects and double identification of the target.

Quantitative assessment of human donor corneal endothelium with Gabor domain optical coherence microscopy.

.We report on a pathway for Gabor domain optical coherence microscopy (GD-OCM)-based metrology to assess the donor’s corneal endothelial layers ex vivo. Six corneas from the Lions Eye Bank at Albany and Rochester were imaged with GD-OCM. The raw 3-D images of the curved corneas were flattened using custom software to enhance the 2-D visualization of endothelial cells (ECs); then the ECs within a circle of -diameter were analyzed using a custom corner method and a cell counting plugin in ImageJ. The EC number, EC area, endothelial cell density (ECD), and polymegethism (CV) were quantified in five different locations for each cornea. The robustness of the method (defined as the repeatability of measurement together with interoperator variability) was evaluated by independently repeating the entire ECD measurement procedure six times by three different examiners. The results from the six corneas show that the current modality reproduces the ECDs with a standard deviation of 2.3% of the mean ECD in every location, whereas the mean ECD across five locations varies by 5.1%. The resolution and imaging area provided through the use of GD-OCM may help to ultimately better assess the quality of donor corneas in transplantation.

Ten Years of Gabor-Domain Optical Coherence Microscopy.

Gabor-domain optical coherence microscopy (GDOCM) is a high-definition imaging technique leveraging principles of low-coherence interferometry, liquid lens technology, high-speed imaging, and precision scanning. GDOCM achieves isotropic 2 μm resolution in 3D, effectively breaking the cellular resolution limit of optical coherence tomography (OCT). In the ten years since its introduction, GDOCM has been used for cellular imaging in 3D in a number of clinical applications, including dermatology, oncology and ophthalmology, as well as to characterize materials in industrial applications. Future developments will enhance the structural imaging capability of GDOCM by adding functional modalities, such as fluorescence and elastography, by estimating thicknesses on the nano-scale, and by incorporating machine learning techniques.

Capabilities of Gabor-domain optical coherence microscopy for the assessment of corneal disease.

.To identify the microstructural modification of the corneal layers during the course of the disease, optical technologies have been pushing the boundary of innovation to achieve cellular resolution of deep layers of the cornea. Gabor-domain optical coherence microscopy (GD-OCM), an optical coherence tomography-based technique that can achieve an isotropic of resolution over a volume of , was developed to investigate the microstructural modifications of corneal layers in four common corneal diseases. Since individual layer visualization without cutting through several layers is challenging due to corneal curvature, a flattening algorithm was developed to remove the global curvature of the endothelial layer and display the full view of the endothelium and Descemet’s membrane in single en face images. As a result, GD-OCM revealed the qualitative changes in size and reflectivity of keratocytes in Fuchs endothelial corneal dystrophy (FECD), which varied by the degree of disease. More importantly, elongated shape and hyperactivation characteristics of keratocytes, associated with the early development of guttae, appeared to start in the posterior stroma very early in the disease process and move toward the anterior stroma during disease progression. This work opens a venue into the pathogenesis of FECD.

Potential of High resolution Gabor‐Domain optical coherence microscopy for early diagnosis of corneal disease

PurposeWe investigate the capability of a high volumetric‐resolution imaging modality, Gabor‐Domain Optical Coherence Microscopy (GD‐OCM), to identify key features in the structural modification of the cornea in three frequent diseases.MethodsHost corneal buttons were excised during penetrating keratoplasty from patients with Fuchs’ endothelial corneal dystrophy (FECD), type IIIA lattice corneal dystrophy (LCD) and keratoconus. We sutured them onto cadaver corneal rims to preserve their shape, mounted onto a corneal artificial chamber and pressurized under a pressure of 20 mmHg. We imaged them using GD‐OCM in a non‐contact mode, combining optical coherence tomography's high sectioning capability with confocal microscopy's high lateral resolution. The system achieved high‐contrast imaging with a field of view of 1 × 1 mm2 and volumetric cellular resolution of 2 μm across a tissue up to 2 mm thick.ResultsGD‐OCM produced high resolution and high‐contrast 3D images of the cornea's different layers. For FECD: the Descemet's membrane and stroma thickened and the density and size of keratocytes increased, along with associated guttae. Change in keratocytes seems to start in the posterior stroma in the early disease process and later heads toward the anterior stroma. For LCD: lattice linear deposits were identified in the anterior stroma, and for keratoconus, folds were identified in the posterior stroma, presumably Vogt's striae. For both, keratocytes and endothelial cells morphology was preserved.ConclusionsThe GD‐OCM revealed key pathologic features of several important corneal diseases and can be applied toward studying corneal diseases.

Optical Assessment of Soft Contact Lens Edge-Thickness.

To assess the edge shape of soft contact lenses using Gabor-Domain Optical Coherence Microscopy (GD-OCM) with a 2-μm imaging resolution in three dimensions and to generate edge-thickness profiles at different distances from the edge tip of soft contact lenses. A high-speed custom-designed GD-OCM system was used to produce 3D images of the edge of an experimental soft contact lens (Bausch + Lomb, Rochester, NY) in four different configurations: in air, submerged into water, submerged into saline with contrast agent, and placed onto the cornea of a porcine eyeball. An algorithm to compute the edge-thickness was developed and applied to cross-sectional images. The proposed algorithm includes the accurate detection of the interfaces between the lens and the environment, and the correction of the refraction error. The sharply defined edge tip of a soft contact lens was visualized in 3D. Results showed precise thickness measurement of the contact lens edge profile. Fifty cross-sectional image frames for each configuration were used to test the robustness of the algorithm in evaluating the edge-thickness at any distance from the edge tip. The precision of the measurements was less than 0.2 μm. The results confirmed the ability of GD-OCM to provide high-definition images of soft contact lens edges. As a nondestructive, precise, and fast metrology tool for soft contact lens measurement, the integration of GD-OCM in the design and manufacturing of contact lenses will be beneficial for further improvement in edge design and quality control. In the clinical perspective, the in vivo evaluation of the lens fitted onto the cornea will advance our understanding of how the edge interacts with the ocular surface. The latter will provide insights into the impact of long-term use of contact lenses on the visual performance.

MEMS-based handheld scanning probe with pre-shaped input signals for distortion-free images in Gabor-domain optical coherence microscopy.

High-speed scanning in optical coherence tomography (OCT) often comes with either compromises in image quality, the requirement for post-processing of the acquired images, or both. We report on distortion-free OCT volumetric imaging with a dual-axis micro-electro-mechanical system (MEMS)-based handheld imaging probe. In the context of an imaging probe with optics located between the 2D MEMS and the sample, we report in this paper on how pre-shaped open-loop input signals with tailored non-linear parts were implemented in a custom control board and, unlike the sinusoidal signals typically used for MEMS, achieved real-time distortion-free imaging without post-processing. The MEMS mirror was integrated into a compact, lightweight handheld probe. The MEMS scanner achieved a 12-fold reduction in volume and 17-fold reduction in weight over a previous dual-mirror galvanometer-based scanner. Distortion-free imaging with no post-processing with a Gabor-domain optical coherence microscope (GD-OCM) with 2 μm axial and lateral resolutions over a field of view of 1 × 1 mm2 is demonstrated experimentally through volumetric images of a regular microscopic structure, an excised human cornea, and in vivo human skin.

Spectral fusing Gabor domain optical coherence microscopy.

Gabor domain optical coherence microscopy (GD-OCM) is one of many variations of optical coherence tomography (OCT) techniques that aims for invariant high resolution across a 3D field of view by utilizing the ability to dynamically refocus the imaging optics in the sample arm. GD-OCM acquires multiple cross-sectional images at different focus positions of the objective lens, and then fuses them to obtain an invariant high-resolution 3D image of the sample, which comes with the intrinsic drawback of a longer processing time as compared to conventional Fourier domain OCT. Here, we report on an alternative Gabor fusing algorithm, the spectral-fusion technique, which directly processes each acquired spectrum and combines them prior to the Fourier transformation to obtain a depth profile. The implementation of the spectral-fusion algorithm is presented and its performance is compared to that of the prior GD-OCM spatial-fusion approach. The spectral-fusion approach shows twice the speed of the spatial-fusion approach for a spectrum size of less than 2000 point sampling, which is a commonly used spectrum size in OCT imaging, including GD-OCM.

Assessing microstructures of the cornea with Gabor-domain optical coherence microscopy: pathway for corneal physiology and diseases.

Gabor-domain optical coherence microscopy (GD-OCM) was applied ex vivo in the investigation of corneal cells and their surrounding microstructures with particular attention to the corneal endothelium. Experiments using fresh pig eyeballs, excised human corneal buttons from patients with Fuchs' endothelial dystrophy (FED), and healthy donor corneas were conducted. Results show in a large field of view (1 mm×1 mm) high definition images of the different cell types and their surrounding microstructures through the full corneal thickness at both the central and peripheral locations of porcine corneas. Particularly, an image of the endothelial cells lining the bottom of the cornea is highlighted. As compared to healthy human corneas, the corneas of individuals with FED show characteristic microstructural alterations of the Descemet's membrane and increased size and number of keratocytes. The GD-OCM-based imaging system developed may constitute a novel tool for corneal imaging and disease diagnosis. Also, importantly, it may provide insights into the mechanism of corneal physiology and pathology, particularly in diseases of the corneal endothelium.

Parallelized multi–graphics processing unit framework for high-speed Gabor-domain optical coherence microscopy

Gabor-domain optical coherence microscopy (GD-OCM) is a volumetric high-resolution technique capable of acquiring three-dimensional (3-D) skin images with histological resolution. Real-time image processing is needed to enable GD-OCM imaging in a clinical setting. We present a parallelized and scalable multi-graphics processing unit (GPU) computing framework for real-time GD-OCM image processing. A parallelized control mechanism was developed to individually assign computation tasks to each of the GPUs. For each GPU, the optimal number of amplitude-scans (A-scans) to be processed in parallel was selected to maximize GPU memory usage and core throughput. We investigated five computing architectures for computational speed-up in processing 1000×1000 A-scans. The proposed parallelized multi-GPU computing framework enables processing at a computational speed faster than the GD-OCM image acquisition, thereby facilitating high-speed GD-OCM imaging in a clinical setting. Using two parallelized GPUs, the image processing of a 1×1×0.6 mm3 skin sample was performed in about 13 s, and the performance was benchmarked at 6.5 s with four GPUs. This work thus demonstrates that 3-D GD-OCM data may be displayed in real-time to the examiner using parallelized GPU processing.

Nondestructive metrology by optical coherence tomography empowering manufacturing iterations of layered polymeric optical materials

In recent years, there has been an ever-growing interest in exploring different optical materials and components to develop compact and effective optical systems. The design and fabrication of high-performance optics require nondestructive metrology techniques to inspect the samples. We have investigated the capability of optical coherence tomography (OCT) to nondestructively characterize layered polymeric materials. Using a custom developed Gabor-domain optical coherence microscopy system centered at 800 nm with 120 nm full width at half maximum enabling unprecedented 2 μm resolution both laterally and axially in an 8 mm 3 volume, we investigated the internal structure of 50 μm thick films and layered sheets, which prompted the manufacturing process to adopt a compatibilization technique. Based on a custom swept-source OCT system centered at 1320 nm with expanded imaging field-of-view and latest depth of imaging extended to ∼5 mm , we performed nondestructive metrology of the layer thickness profiles over the depth of a monolithic layered sheet and diagnosed a film compression issue within the sheet. With the OCT metrology, the manufacturing process has been advanced and the layer thickness profile of a recent layered gradient refractive index sheet shows improved uniformity through depth.

Three-dimensional imaging of normal skin and nonmelanoma skin cancer with cellular resolution using Gabor domain optical coherence microscopy

We investigate morphological differences in three-dimensional (3-D) images with cellular resolution between nonmelanoma skin cancer and normal skin using Gabor domain optical coherence microscopy. As a result, we show for the first time cellular optical coherence images of 3-D features differentiating cancerous skin from normal skin. In addition, in vivo volumetric images of normal skin from different anatomic locations are shown and compared.

Assessment of a liquid lens enabled in vivooptical coherence microscope

The optical aberrations induced by imaging through skin can be predicted using formulas for Seidel aberrations of a plane-parallel plate. Knowledge of these aberrations helps to guide the choice of numerical aperture (NA) of the optics we can use in an implementation of Gabor domain optical coherence microscopy (GD-OCM), where the focus is the only aberration adjustment made through depth. On this basis, a custom-designed, liquid-lens enabled dynamic focusing optical coherence microscope operating at 0.2 NA is analyzed and validated experimentally. As part of the analysis, we show that the full width at half-maximum metric, as a characteristic descriptor for the point spread function, while commonly used, is not a useful metric for quantifying resolution in non-diffraction-limited systems. Modulation transfer function (MTF) measurements quantify that the liquid lens performance is as predicted by design, even when accounting for the effect of gravity. MTF measurements in a skinlike scattering medium also quantify the performance of the microscope in its potential applications. To guide the fusion of images across the various focus positions of the microscope, as required in GD-OCM, we present depth of focus measurements that can be used to determine the effective number of focusing zones required for a given goal resolution. Subcellular resolution in an onion sample, and high-definition in vivo imaging in human skin are demonstrated with the custom-designed and built microscope.