Full-field Optical Coherence Tomography Images Research Articles

Influence of static and dynamic ocular aberrations on full-field optical coherence tomography for in vivo high-resolution retinal imaging.

Under spatially incoherent illumination, time-domain full-field optical coherence tomography (FFOCT) offers the possibility to achieve in vivo retinal imaging at cellular resolution over a wide field of view. Such performance is possible, albeit there is the presence of ocular aberrations even without the use of classical adaptive optics. While the effect of aberrations in FFOCT has been debated these past years, mostly on low-order and static aberrations, we present, for the first time to our knowledge, a method enabling a quantitative study of the effect of statistically representative static and dynamic ocular aberrations on FFOCT image metrics, such as SNR, resolution, and image similarity. While we show that ocular aberrations can decrease FFOCT SNR and resolution by up to 14 dB and fivefold, we take advantage of such quantification to discuss different possible compromises between performance gain and adaptive optics complexity and speed, to optimize both sensor-based and sensorless FFOCT high-resolution retinal imaging.

Training With Uncertain Annotations for Semantic Segmentation of Basal Cell Carcinoma From Full-Field OCT Images.

Semantic segmentation of basal cell carcinoma (BCC) from full-field optical coherence tomography (FF-OCT) images of human skin has received considerable attention in medical imaging. However, it is challenging for dermatopathologists to annotate the training data due to OCT's lack of color specificity. Very often, they are uncertain about the correctness of the annotations they made. In practice, annotations fraught with uncertainty profoundly impact the effectiveness of model training and hence the performance of BCC segmentation. To address this issue, we propose an approach to model training with uncertain annotations. The proposed approach includes a data selection strategy to mitigate the uncertainty of training data, a class expansion to consider sebaceous gland and hair follicle as additional classes to enhance the performance of BCC segmentation, and a self-supervised pre-training procedure to improve the initial weights of the segmentation model parameters. Furthermore, we develop three post-processing techniques to reduce the impact of speckle noise and image discontinuities on BCC segmentation. The mean Dice score of BCC of our model reaches 0.503±0.003, which, to the best of our knowledge, is the best performance to date for semantic segmentation of BCC from FF-OCT images.

Single-shot off-axis full-field optical coherence tomography

Full field optical coherence tomography (FF-OCT) enables high-resolution in-depth imaging within turbid media. In this work, we present a simple approach which combines FF-OCT with off-axis interferometry for reconstruction of en-face images. With low spatial and temporal coherence illumination, this method is able to extract an FF-OCT image from only one interference acquisition. This method is described, and the proof-of-concept is demonstrated through the observation of scattering samples such as organic and ex vivo biomedical samples.

Photoaging and Sequential Function Reversal with Cellular-Resolution Optical Coherence Tomography in a Nude Mice Model.

Although nude mice are an ideal photoaging research model, skin biopsies result in inflammation and are rarely performed at baseline. Meanwhile, studies on antiphotoaging antioxidants or rejuvenation techniques often neglect the spontaneous reversal capacity. Full-field optical coherence tomography (FFOCT) can acquire cellular details noninvasively. This study aimed to establish a photoaging and sequential function reversal nude mice model assisted by an in vivo cellular resolution FFOCT system. We investigated whether a picosecond alexandrite laser (PAL) with a diffractive lens array (DLA) accelerated the reversal. In the sequential noninvasive assessment using FFOCT, a spectrophotometer, and DermaLab Combo®, the photodamage percentage recovery plot demonstrated the spontaneous recovery capacity of the affected skin by UVB-induced transepidermal water loss and UVA-induced epidermis thickening. A PAL with DLA not only accelerated skin barrier regeneration with epidermal polarity, but also increased dermal neocollagenesis, whereas the nonlasered group still had >60% collagen intensity loss and 40% erythema from photodamage. Our study demonstrated that FFOCT images accurately resemble the living tissue. The photoaging and sequential function reversal model provides a reference to assess the spontaneous recovery capacity of nude mice from photodamage. This model can be utilized to evaluate the sequential noninvasive photodamage and reversal effects after other interventions.

Optimization of the performance of hand‐held FFOCT scanner

In this work, we have established an improved handheld full-field optical coherence tomography (FFOCT) system based on the tandem structure of Michelson interferometer and Fizeau interferometer. We have analyzed the influence of wave aberrations of the handheld FFOCT as a function of orientation deviation of mirror M2 on the system point spread function (PSF) by using the nodal aberration theory. It is found that the range of the orientation deviation of mirror M2 must be less than 0.1° to generate stable FFOCT images. The effect of wave aberrations of defocus as a result of the refractive index difference on the system PSF has also been calculated, the measuring depth in the case of the sample with refractive index of 1.5 and the propagation medium is air (n = 1) should be less than 20 μm. Theoretical analyses have been demonstrated by the experimental results of aluminum powder, onion and porcine myocardium. By comparing the defocused FFOCT images of aluminum powder with defocused FFOCT images of onion and porcine myocardium, it can be concluded that reducing the refractive index difference between the medium and sample can increase the imaging depth of handheld FFOCT system.

Computer-Aided Detection (CADe) System with Optical Coherent Tomography for Melanin Morphology Quantification in Melasma Patients.

Dark skin-type individuals have a greater tendency to have pigmentary disorders, among which melasma is especially refractory to treat and often recurs. Objective measurement of melanin amount helps evaluate the treatment response of pigmentary disorders. However, naked-eye evaluation is subjective to weariness and bias. We used a cellular resolution full-field optical coherence tomography (FF-OCT) to assess melanin features of melasma lesions and perilesional skin on the cheeks of eight Asian patients. A computer-aided detection (CADe) system is proposed to mark and quantify melanin. This system combines spatial compounding-based denoising convolutional neural networks (SC-DnCNN), and through image processing techniques, various types of melanin features, including area, distribution, intensity, and shape, can be extracted. Through evaluations of the image differences between the lesion and perilesional skin, a distribution-based feature of confetti melanin without layering, two distribution-based features of confetti melanin in stratum spinosum, and a distribution-based feature of grain melanin at the dermal–epidermal junction, statistically significant findings were achieved (p-values = 0.0402, 0.0032, 0.0312, and 0.0426, respectively). FF-OCT enables the real-time observation of melanin features, and the CADe system with SC-DnCNN was a precise and objective tool with which to interpret the area, distribution, intensity, and shape of melanin on FF-OCT images.

Effects of orientation deviation of a beam splitter and a reference mirror on the stability of a two-interferometer-based handheld FFOCT imaging probe

In this work, we present a handheld full-field optical coherence tomography (FFOCT) system based on a series connection of two interferometers: the Michelson interferometer is used as a compensation part and the Fizeau interferometer is used as a detection part. Owing to the common-path arrangement of the Fizeau interferometer, this handheld FFOCT system has a compact detection arm and is insensitive to the external disturbance. A high-output halogen lamp and high NA microscope objective contribute to achieving the spatial resolution of 0.7µm×0.5µm (transverse × axial). Low imaging stability is caused by an extremely short coherence length. We found that to generate en face images with high quality and high stability using a probe with an extremely short coherence length, the range of deviation of the orientation of the beam splitter must be less than 1°, and the range of orientation deviation of the mirror in the Michelson interferometer corresponds to the displacement between the two field stop images at a distance not to exceed 10 µm.

Nonlocally adaptive image enhancement system for full-field optical coherence tomography

Full-field optical coherence tomography (FFOCT) can be used to acquire cellular-level morphological images of nonfixed and unstained biological tissues. However, the speckle noise and vignetting of FFOCT images are drawbacks in terms of pathological application. We present a novel adaptive nonlocal image enhancement algorithm that uses a fluorescence microscopy (FM) image of the nucleus to achieve cytoplasm image enhancement under FFOCT. The algorithm comprises a denoising module and an adaptive nonlocal enhancement module. The denoising module is an end-to-end residual network with skip connections and dilated convolution. The adaptive nonlocal enhancement module enhances images to segment tissues, nontissues, and transition areas so that the images retain the details of the original cell structure but do not contain noise. The proposed algorithm was implemented on a dual-modality scanner system that combines FFOCT and FM. A total of 12 pig liver and pig kidney tissues were sectioned into 10,236 samples for experiments. The peak signal-to-noise ratio and structural similarity index value increased from 30.8 and 0.58, respectively, for the aforementioned system without the proposed algorithm to 34.2 and 0.86, respectively, for the aforementioned system with the proposed algorithm. Ten samples of human breast tissues were used to validate the proposed algorithm. The average contrast-to-noise ratio was 1.69 and 11.73 when the aforementioned system was used with and without the proposed algorithm, respectively. The results of this study revealed that the quality of scanned images considerably improved when the proposed system was used and that the proposed system can facilitate clinical pathological diagnosis.

Simultaneous 4-phase-shifted full-field optical coherence microscopy.

A new method is presented for full-field optical coherence tomography imaging, which permits capturing single shot phase sensitive imaging through simultaneous acquisition of four phase-shifted images with a single camera using unpolarized light for object illumination. Our method retains the full dynamic range of the camera by using different areas of a single camera sensor to capture each image. We demonstrate the performance of our method by imaging phantoms and live cultures of fibroblast, cancer, and macrophage cells to achieve 59 dB sensitivity with isotropic resolution down to 1 μm, and displacement sensitivity down to 0.1 nm. Our method can serve as a platform for developing high resolution imaging systems because when used in conjunction with broadband spatially incoherent light sources, the resolution is not affected by optical aberrations or speckle noise.

Analysis of the impact of optical aberrations in en-face full-field OCT microscopy.

Optical coherence tomography (OCT) is a powerful technique for cross-sectioning imaging. However, the lateral resolution may be degraded by optical aberrations originating from the sample or the setup. We present an extensive quantitative study of the impact of aberrations in time-domain en-face full-field OCT (FFOCT). Using an adaptive optics loop integrated in an FFOCT setup, a deformable mirror is used to introduce low-order calibrated aberrations. The experimental analysis of both the line spread functions (SF) and the complex object images has allowed us to measure the loss in contrast and the impact on lateral spatial resolution. We demonstrate that the frequency content of FFOCT image spectra in terms of signal-to-noise ratio and cutoff frequency is degraded by aberrations but remains much higher than in conventional incoherent images. Line SF profiles in conventional imaging display widening, whereas in FFOCT they display oscillations, leading to the possible perception of preserved resolution. Nevertheless, for complex objects, the aberration image blurring is strong due to the convolution process by the point SF, resulting in a significant filtering of the image spatial spectrum.

Detecting mouse squamous cell carcinoma from submicron full-field optical coherence tomography images by deep learning.

The standard medical practice for cancer diagnosis requires histopathology, which is an invasive and time-consuming procedure. Optical coherence tomography (OCT) is an alternative that is relatively fast, noninvasive, and able to capture three-dimensional structures of epithelial tissue. Unlike most previous OCT systems, which cannot capture crucial cellular-level information for squamous cell carcinoma (SCC) diagnosis, the full-field OCT (FF-OCT) technology used in this paper is able to produce images at sub-micron resolution and thereby facilitates the development of a deep learning algorithm for SCC detection. Experimental results show that the SCC detection algorithm can achieve a classification accuracy of 80% for mouse skin. Using the sub-micron FF-OCT imaging system, the proposed SCC detection algorithm has the potential for in-vivo applications.

Single-shot surface 3D imaging by optical coherence factor.

We report a single-shot three-dimensional (3D) topographical imaging method, optical coherence factor (OCF) imaging, which uses optical coherence as the contrast mechanism to acquire the surface height (${z}$z-direction) information of an object. A 4-f imaging system records the light field reflected from the surface of the object. The illumination of the imaging system comes from a laser source with the optical coherence length comparable to the depth of field (DoF) of the optical system. Off-axis holographic recording is used to retrieve the coherence factor from the interference fringes, which is then converted to ${z}$z-direction information. In this experiment, we validate our 3D imaging results comparing them to axial scanning full-field optical coherence tomography images. We also analyze the contrast mechanism of OCF and show that it is able to provide additional information over conventional coherent and incoherent imaging using the same imaging setup. This single-shot computationally efficient method may have potential applications in industrial quality control inspection.

Liver tissue classification of en face images by fractal dimension-based support vector machine.

Full-field optical coherence tomography (FF-OCT) has been reported with its label-free subcellular imaging performance. To realize quantitive cancer detection, the support vector machine model of classifying normal and cancerous human liver tissue is proposed with en face tomographic images. Twenty samples (10 normal and 10 cancerous) were operated from humans and composed of 285 en face tomographic images. Six histogram features and one proposed fractal dimension parameter that reveal the refractive index inhomogeneities of tissue were extracted and made up the training set. The other different 16 samples (8 normal and 8 cancerous) were imaged (190 images) and employed as the test set with the same features. First, a subcellular-resolution tomographic image library for four histopathological areas in liver tissue was established. Second, the area under the receiver operating characteristics of 0.9378, 0.9858, 0.9391, 0.9517 for prediction of the cancerous hepatic cell, central vein, fibrosis, and portal vein were measured with the test set. The results indicate that the proposed classifier from FF-OCT images shows promise as a label-free assessment of quantified tumor detection, suggesting the fractal dimension-based classifier could aid clinicians in detecting tumor boundaries for resection in surgery in the future.

Comparison of full-field optical coherence tomography imaging for pancreatic tissue sample obtained by EUS-fine-needle biopsy and conventional histological examination: A study protocol for a prospective trial.

For a definitive diagnosis of fine-needle aspiration (FNA)/biopsy, one of the reliable techniques to determine the adequacy and accuracy rapid on-site evaluation (ROSE) of cytological samples is preferable. Because of the lack of trained pathologists, alternatives have to be explored. This study is primarily conducted to determine the diagnostic sensitivity and specificity of full-field optical coherence tomography (FF-OCT) and secondarily to evaluate the possibility of FF-OCT differentiating different types of pancreatic diseases. The diagnostic coherence of FF-OCT by a trained assistant (endoscopist) and trained pathologist is also compared. This is a single-center, prospective, observation trial. Eighty patients would be enrolled in the study. The tissue samples acquired by endoscopic ultrasound fine-needle biopsy (EUS-FNB) would be imaged by the FF-OCT system, interpreted by a trained endoscopist and a pathologist. The results of the image interpretation would be verified with histological findings. This study determines the diagnostic capability of FF-OCT as a ROSE technique while performing EUS-FNB, and whether endoscopists can implement the assessment.

FFOCT imaging based on compressed sensing

With the demand for high-resolution images, the storage space occupied by the images of the full-field optical coherence tomography (FFOCT) system is becoming more and more intensified. How to reduce the data volume of high-resolution image systems has become a research hotspot in the image processing industry. In particular, precision medicine requires high-resolution images of organs and tissues, and this data storage system requires a large storage space. For this reason, based on the high-resolution tomographic image obtained from our organ tissue, compressed sensing theory is used to perform compression reconstruction simulation. It is verified that the amount of data stored in the three-dimensional image of the diseased tissue is effectively reduced without changing the image resolution.

Modeling of full-field optical coherence tomography in scattering media.

We develop a model of full-field optical coherence tomography (FF-OCT) that includes a description of partial temporal and spatial coherence, together with a mean-field scattering theory going beyond the Born approximation. Based on explicit expressions of the FF-OCT signal, we discuss essential features of FF-OCT imaging, such as the influence of partial coherence on the optical transfer function, and on the decay of the signal with depth. We derive the conditions under which the spatially averaged signal exhibits a pure exponential decay, providing a clear frame for the use of the Beer-Lambert law for quantitative measurements of the extinction length in scattering media.

Image contrast correction method in full-field optical coherence tomography

An image contrast correction method is proposed for en face images obtained with full-field optical coherence tomography (FFOCT). First, the mechanism of image contrast decrease in FFOCT is considered and the theoretical models of main parameters that degrade image contrast are analyzed. Second, changes of contrast with depth are calculated under various conditions, from which the main parameters that affect contrast in tissue are identified. Then based on the analysis, the methods are proposed for correcting image contrast. Finally, the en face tomographic images of human liver tissue at different depths with and without contrast correction are presented to demonstrate the capability of the method. The results are very helpful for correctly interpreting FFOCT images for applications in disease diagnosis.

Qualitative and quantitative assessment of cartilage degeneration using full-field optical coherence tomography exvivo.

The purpose of this study was to investigate the ability of full-field optical coherence tomography (FFOCT) to qualitatively and quantitatively evaluate cartilage degeneration using the qualitative evaluation of histology sections as the reference. Thirty-three human knee cartilage samples of variable degeneration were included in the study. A closely matching histology and FFOCT image was acquired for each sample. The cartilage degeneration was qualitatively evaluated by assigning a grade to each histology and FFOCT image. The relevance of the performed grading was assessed by calculating the intra- and inter-observer reproducibility and calculating the concordance between the histology and FFOCT grades. A near-automatic algorithm was developed to quantitatively characterize the cartilage surface in each image. The correlation between the quantitative results and the reference qualitative histology was calculated. An almost perfect agreement was achieved for both the intra- and inter-reproducibility of the histology and FFOCT qualitative grading (κ≥0.91). A high and statistically significant level of agreement was measured between the histology and FFOCT grades (W=0.95, P<0.05). Strong and statistically significant correlations were measured between the quantitative results and the reference qualitative histology grades (ρ≥0.75, P<0.05). We have demonstrated that FFOCT is an alternative approach to conventional optical coherence tomography (OCT) that is as well adapted for the qualitative and quantitative assessment of human cartilage as the reference gold standard - histology. This study constitutes the first promising results towards developing a new diagnostic tool in the field of osteoarthritis.

Cell Motility as Contrast Agent in Retinal Explant Imaging With Full-Field Optical Coherence Tomography.

To use cell motility as a contrast agent in retinal explants. Macaque and mouse retinal explants were imaged with high resolution full field optical coherence tomography (FFOCT) and dynamic FFOCT, coupled with fluorescence imaging. Static and dynamic FFOCT create complementary contrast from different structures within a cell. When imaging in vitro samples, static FFOCT detects steep refractive index gradients to reveal stationary structures including fibers, vessels, collagen, and cell contours, while dynamic FFOCT emphasizes metabolic activity of moving structures that are mainly intracellular, thus creating or enhancing contrast in cells that were previously hidden in noise. Dynamic FFOCT enables detection of most of the retinal cell types in the ganglion cell, inner and outer nuclear layers, where static FFOCT contrast is too low in relation to the noise background. Composite static and dynamic FFOCT provides a new kind of FFOCT image containing valuable information for imaging of retinal explants. It provides label-free en face images of living retinas, with a subcellular resolution. Dynamic FFOCT adds information about cell activity, which is of interest in longitudinal explant studies.

Freeze artifact on full-field optical coherence tomography skin imaging.

Full-field optical coherence tomography (FFOCT) is an emerging imaging technique for rapid histological analysis. As FFOCT is introduced into the Mohs workflow, it is important to document the effect of tissue freezing on FFOCT images and any effect FFOCT has on frozen sections. Our study aimed to evaluate the changes on FFOCT images after tissue freezing as well as FFOCT imaging effects on frozen sectioning. Six normal skin specimens were imaged using FFOCT and subsequently frozen using a cryostat. The specimens were then reimaged using FFOCT and compared for any differences. To evaluate the effect of FFOCT imaging on frozen sections, five normal skin specimens were bisected and one half was imaged using FFOCT. Both halves underwent frozen sectioning and analysis. Significant changes in the fat, sebaceous glands, eccrine glands, and dermal collage and minimal changes in the epidermis were seen after freezing. No effect on frozen sectioning was seen after FFOCT imaging. As FFOCT is studied for use in Mohs, clinicians should be aware that freezing prior to imaging introduces significant artifact in the FFOCT image. If possible, specimens should undergo imaging prior to being frozen.