A compact high‐speed full‐field optical coherence microscope for high‐resolution in vivo skin imaging

A compact high‐speed full‐field optical coherence microscope has been developed for high‐resolution in vivo imaging of biological tissues. The interferometer, in the Linnik configuration, has a size of 11 × 11 × 5 cm³ and a weight of 210 g. Full‐field illumination with low‐coherence light is achieved with a high‐brightness broadband light‐emitting diode. High‐speed full‐field detection is achieved by using part of the image sensor of a high‐dynamic range CMOS camera. En face tomographic images are acquired at a rate of 50 Hz, with an integration time of 0.9 ms. The image spatial resolution is 0.9 μm × 1.2 μm (axial × transverse), over a field of view of 245 × 245 μm². Images of human skin, revealing in‐depth cellular‐level structures, were obtained in vivo and in real‐time without the need for stabilization of the subject. The system can image larger fields, up to 1 × 1 mm², but at a reduced depth.      

Phase unwrapping for Doppler spectral domain optical coherence tomography flow measurement

Doppler optical coherence tomography (OCT) offers additional flow velocity information, which extends the application of OCT. Phase wrapping is the inherent problem that limits measureable range of Doppler OCT. We propose a phase unwrapping method which is suitable for correcting phase in Doppler OCT images. Points (pixels) in flow region are divided into groups according to the radial distance. Points in the same group are supposed to have close velocity. Phase unwrapping algorithm begins at the boundary layer group and is performed sequentially toward the center. Using the proposed criterion, points in a group are separated into two categories, signal points and noise points. Wrapping rounds are determined for signal points phase unwrapping. Mean value of the corrected signal points replaces the noise points for noise reduction. The method is validated with capillary tube flow phantom and in vivo blood flow.     

Heart‐beat‐phase‐coherent Doppler optical coherence tomography for measuring pulsatile ocular blood flow

We introduce a Doppler OCT (DOCT) platform that is fully synchronized with the heart‐beat via a pulse oximeter. The system allows reconstructing heart‐beat‐phase‐coherent quantitative DOCT volumes. The method is to acquire a series of DOCT volumes and to record the pulse in parallel. The heartbeat data is used for triggering the start of each DOCT volume acquisition. The recorded volume series is registered to the level of capillaries using a cross‐volume registration. The information of the pulse phase is used to rearrange the tomograms in time, to obtain a series of phase coherent DOCT volumes over a pulse. We present Doppler angle independent quantitative evaluation of the absolute pulsatile blood flow within individual retinal vessels as well as of the total retinal blood flow over a full heartbeat cycle. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Four‐dimensional imaging of murine subpleural alveoli using high‐speed optical coherence tomography

The investigation of lung dynamics on alveolar scale is crucial for the understanding and treatment of lung diseases, such as acute lung injury and ventilator induced lung injury, and to promote the development of protective ventilation strategies. One approach to this is the establishment of numerical simulations of lung tissue mechanics where detailed knowledge about three‐dimensional alveolar structure changes during the ventilation cycle is required. We suggest four‐dimensional optical coherence tomography (OCT) imaging as a promising modality for visualizing the structural dynamics of single alveoli in subpleural lung tissue with high temporal resolution using a mouse model. A high‐speed OCT setup based on Fourier domain mode locked laser technology facilitated the acquisition of alveolar structures without noticeable motion artifacts at a rate of 17 three‐dimensional stacks per ventilation cycle. The four‐dimensional information, acquired in one single ventilation cycle, allowed calculating the volume‐pressure curve and the alveolar compliance for single alveoli. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Closed loop tracked Doppler optical coherence tomography based heart monitor for the Drosophila melanogaster larvae

This paper presents a novel instrument for biosciences, useful for studies of moving embryos. A dual sequential imaging/measurement channel is assembled via a closed‐loop tracking architecture. The dual channel system can operate in two regimes: (i) single‐point Doppler signal monitoring or (ii) fast 3‐D swept source OCT imaging. The system is demonstrated for characterizing cardiac dynamics in Drosophila melanogaster larva. Closed loop tracking enables long term in vivo monitoring of the larvae heart without anesthetic or physical restraint. Such an instrument can be used to measure subtle variations in the cardiac behavior otherwise obscured by the larvae movements.

A fruit fly larva (top) was continuously tracked for continuous remote monitoring. A heartbeat trace of freely moving larva (bottom) was obtained by a low coherence interferometry based doppler sensing technique.     

Multicontrast endomyocardial imaging by single‐channel high‐resolution cross‐polarization optical coherence tomography

A single‐channel high‐resolution cross‐polarization (CP) optical coherence tomography (OCT) system is presented for multicontrast imaging of human myocardium in one‐shot measurement. The intensity and functional contrasts, including the ratio between the cross‐ and co‐polarization channels as well as the cumulative retardation, are reconstructed from the CP‐OCT readout. By comparing the CP‐OCT results with histological analysis, it is shown that the system can successfully delineate microstructures in the myocardium and differentiate the fibrotic myocardium from normal or ablated myocardium based on the functional contrasts provided by the CP‐OCT system. The feasibility of using A‐line profiles from the 2 orthogonal polarization channels to identify fibrotic myocardium, normal myocardium and ablated lesion is also discussed.      

Improving cerebral microvascular image quality of optical coherence tomography angiography with deep learning-based segmentation

Optical coherence tomography angiography (OCTA) can map the microvascular networks of the cerebral cortices with micrometer resolution and millimeter penetration. However, the high scattering of the skull and the strong noise in the deep imaging region will distort the vasculature projections and decrease the OCTA image quality. Here, we proposed a deep learning-based segmentation method based on a U-Net convolutional neural network to extract the cortical region from the OCT image. The vascular networks were then visualized by three OCTA algorithms. The image quality of the vasculature projections was assessed by two metrics, including the peak signal-to-noise ratio (PSNR) and the contrast-to-noise ratio (CNR). The results show the accuracy of the cortical segmentation was 96.07%. The PSNR and CNR values increased significantly in the projections of the selected cortical regions. The OCTA incorporating the deep learning-based cortical segmentation can efficiently improve the image quality and enhance the vasculature clarity.     

Imaging sebaceous gland using optical coherence tomography with deep learning assisted automatic identification

Imaging sebaceous glands and evaluating morphometric parameters are important for diagnosis and treatment of serum problems. In this article, we investigate the feasibility of high-resolution optical coherence tomography (OCT) in combination with deep learning assisted automatic identification for these purposes. Specifically, with a spatial resolution of 2.3 μm × 6.2 μm (axial × lateral, in air), OCT is capable of clearly differentiating sebaceous gland from other skin structures and resolving the sebocyte layer. In order to achieve efficient and timely imaging analysis, a deep learning approach built upon ResNet18 is developed to automatically classify OCT images (with/without sebaceous gland), with a classification accuracy of 97.9%. Based on the result of automatic identification, we further demonstrate the possibility to measure gland size, sebocyte layer thickness and gland density.     

Cross‐scanning optical coherence tomography angiography for eye motion correction

We propose a cross‐scanning optical coherence tomography (CS‐OCT) system to correct eye motion artifacts in OCT angiography images. This system employs a dual‐illumination configuration with two orthogonally polarized beams, each of which simultaneously perform raster scanning in perpendicular direction with each other over the same area. In the reference arm, a polarization delay unit is used to acquire the two orthogonally polarized interferograms with a single photo detector by introducing different optical delay lines. The two cross‐scanned volume data are affected by the same eye motion but in two orthogonal directions. We developed a motion correction algorithm, which removes artifacts in the slow axis of each angiogram using the other and merges them through a nonrigid registration algorithm. In this manner, we obtained a motion‐corrected angiogram within a single volume scanning time without additional eye‐tracking devices.     

Origin of improved depth penetration in dual‐axis optical coherence tomography: a Monte Carlo study

Recent studies have demonstrated that extended imaging depth can be achieved using dual‐axis optical coherence tomography (DA‐OCT). By illuminating and collecting at an oblique angle, multiple forward scattered photons from large probing depths are preferentially detected. However, the mechanism behind the enhancement of imaging depth needs further illumination. Here, the signal of a DA‐OCT system is studied using a Monte Carlo simulation. We modeled light transport in tissue and recorded the spatial and angular distribution of photons exiting the tissue surface. Results indicate that the spatial separation and offset angle created by the non‐telecentric scanning configuration promote the collection of more deeply propagating photons than conventional on‐axis OCT.      

Investigation of the human tympanic membrane oscillation ex vivo by Doppler optical coherence tomography

Investigations of the tympanic membrane (TM) can have an important impact on understanding the sound conduction in the ear and can therefore support the diagnosis and treatment of diseases in the middle ear. High‐speed Doppler optical coherence tomography (OCT) has the potential to describe the oscillatory behaviour of the TM surface in a phase‐sensitive manner and additionally allows acquiring a three‐dimensional image of the underlying structure. With repeated sound stimuli from 0.4 kHz to 6.4 kHz, the whole TM can be set in vibration and the spatially resolved frequency response functions (FRFs) of the tympanic membrane can be recorded. Typical points, such as the umbo or the manubrium of malleus, can be studied separately as well as the TM surface with all stationary and wave‐like vibrations. Thus, the OCT methodology can be a promising technique to distinguish between normal and pathological TMs and support the differentiation between ossicular and membrane diseases. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

K‐distribution three‐dimensional mapping of biological tissues in optical coherence tomography

Probability density function (PDF) analysis with K‐distribution model of optical coherence tomography (OCT) intensity signals has previously yielded a good representation of the average number of scatterers in a coherence volume for microspheres‐in‐water systems, and has shown initial promise for biological tissue characterization. In this work, we extend these previous findings, based on single point M‐mode or two‐dimenstional slice analysis, to full three‐dimensional (3D) imaging maps of the shape parameter α of the K‐distribution PDF. After selecting a suitably sized 3D evaluation window, and verifying methodology in phantoms, the resultant parametric α images obtained in different animal tissues (rat liver and brain) show new contrasting ability not seen in conventional OCT intensity images.      

Interplane bulk motion analysis and removal based on normalized cross‐correlation in optical coherence tomography angiography

Bulk motion seriously degrades the image quality of optical coherence tomography angiography (OCTA). Conventional correction methods focus on in‐plane displacement, while the bulk motion component perpendicular to B‐scans also introduces noise. This work first presents an evaluation of this component using a specific scan protocol and an approximate expression derived from peak‐normalized cross‐correlation values, and then quantitatively assesses how interplane bulk motion noise reduce the sensitivity of cross‐sectional angiograms. Finally, we developed a repetitive bulk motion correction method based on the estimated displacements and redundant volume scans. The correction does not require registration and angiogram reconstruction of low flow sensitivity frames, and the results of in vivo mice skin OCTA imaging experiments show that the proposed method can effectively reduce bulk motion noise caused by cardiac and respiratory motion and occasional shaking, and improve OCTA image quality, which has practical significance for clinical OCTA diagnosis and analysis.     

Polarization‐multiplexed,dual‐beam swept source optical coherence tomography angiography

A polarization‐multiplexed, dual‐beam setup is proposed to expand the field of view (FOV) for a swept source optical coherence tomography angiography (OCTA) system. This method used a Wollaston prism to split sample path light into 2 orthogonal‐polarized beams. This allowed 2 beams to shine on the cornea at an angle separation of ~14°, 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 2 polarized beams so the interferogram from the 2 beams are coded at different frequency bands. The resulting OCTA images from the 2 beams were coded with a depth separation of ~2 mm. A total of 5 × 5 mm² angiograms from the 2 beams were obtained simultaneously in 4 seconds. The 2 angiograms then were montaged to get a wider FOV of ~5 × 9.2 mm².      

Three‐dimensional multiphoton/optical coherence tomography for diagnostic applications in dermatology

A preliminary clinical trial using state‐of‐the‐art multiphoton tomography (MPT) and optical coherence tomography (OCT) for three‐dimensional (3D) multimodal in vivo imaging of normal skin, nevi, scars and pathologic skin lesions has been conducted. MPT enabled visualization of sub‐cellular details with axial and transverse resolutions of <2 μm and <0.5 μm, respectively, from a volume of 0.35 × 0.35 × 0.2 mm³ at a frame rate of 0.14 Hz (512 × 512 pixels). State‐of‐the‐art OCT, operating at a center wavelength of 1300 nm, was capable of acquiring 3D images depicting the layered architecture of skin with axial and transverse resolutions ～8 μm and ～20 μm, respectively, from a volume of 7 × 3.5 × 1.5 mm³ at a frame rate of 46 Hz (1024 × 1024 pixels). This study demonstrates the clinical diagnostic potential of MPT/OCT for pre‐screening relatively large areas of skin using 3D OCT to identify suspicious regions at microscopic level and subsequently using high resolution MPT to obtain zoomed in, sub‐cellular level information of the respective regions (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Measuring polarization changes in the human outer retina with polarization‐sensitive optical coherence tomography

Morphological changes in the outer retina such as drusen are established biomarkers to diagnose age‐related macular degeneration. However, earlier diagnosis might be possible by taking advantage of more subtle changes that accompany tissues that bear polarization‐altering properties. To test this hypothesis, we developed a method based on polarization‐sensitive optical coherence tomography with which volumetric data sets of the macula were obtained from 10 young (<25 years) and 10 older (>54 years) subjects. All young subjects and 5 of the older subjects had retardance values induced by the retinal pigment epithelium and Bruch's membrane (RPE‐BM) complex that were just above the noise floor measurement (5°‐13° at 840 nm). In contrast, elevated retardance, up to 180°, was observed in the other 5 older subjects. Analysis of the degree of polarization uniformity (DOPU) demonstrates that reduced DOPU (<0.4) in the RPE is associated with elevated double pass phase retardation (DPPR) below the RPE‐BM complex, suggesting that the observed elevated DPPR in older subjects is the result of increased scattering or polarization scrambling. Collectively, our measurements show that the outer retina can undergo dramatic change in its polarization properties with age, and in some cases still retain its clinically normal appearance.      

Machine learning in optical coherence tomography angiography

Optical coherence tomography angiography (OCTA) offers a noninvasive label-free solution for imaging retinal vasculatures at the capillary level resolution. In principle, improved resolution implies a better chance to reveal subtle microvascular distortions associated with eye diseases that are asymptomatic in early stages. However, massive screening requires experienced clinicians to manually examine retinal images, which may result in human error and hinder objective screening. Recently, quantitative OCTA features have been developed to standardize and document retinal vascular changes. The feasibility of using quantitative OCTA features for machine learning classification of different retinopathies has been demonstrated. Deep learning-based applications have also been explored for automatic OCTA image analysis and disease classification. In this article, we summarize recent developments of quantitative OCTA features, machine learning image analysis, and classification.     

Full‐field swept‐source optical coherence tomography and neural tissue classification for deep brain imaging

Optical coherence tomography can differentiate brain regions with intrinsic contrast and at a micron scale resolution. Such a device can be particularly useful as a real‐time neurosurgical guidance tool. We present, to our knowledge, the first full‐field swept‐source optical coherence tomography system operating near a wavelength of 1310 nm. The proof‐of‐concept system was integrated with an endoscopic probe tip, which is compatible with deep brain stimulation keyhole neurosurgery. Neuroimaging experiments were performed on ex vivo brain tissues and in vivo in rat brains. Using classification algorithms involving texture features and optical attenuation, images were successfully classified into three brain tissue types.     

Flexible wide‐field optical micro‐angiography based on Fourier‐domain multiplexed dual‐beam swept source optical coherence tomography

Wide‐field optical coherence tomography angiography (OCTA) is gaining interest in clinical imaging applications. In this pursuit, it is challenging to maintain the imaging resolution and sensitivity throughout the wide field of view (FoV). Here, we propose a novel method/system of dual‐beam arrangement and Fourier‐domain multiplexing to achieve wide‐field OCTA when imaging the uneven surface samples. The proposed system provides 2 separate FoVs, with flexibility that the imaging area, focus of the imaging beam and imaging depth range can be individually adjusted for each FoV, leading to either (1) increased system imaging FoV or (2) capability of targeting 2 regions of interests that locate at depths with large difference between each other. We demonstrate this novel method by employing 100 kHz laser source in a swept source OCTA to achieve an effective 200 kHz sweeping rate, covering a 22 × 22 mm FoV. The results are verified by a SS‐OCTA system employing a 200 kHz laser source, together with the experimental demonstrations when imaging whole brain vasculature in rodent models and skin blood perfusion in human fingers, show‐casing the capability of proposed system to image live large samples with complex surface topography.      

Comparative study of optical coherence tomography angiography algorithms for rodent retinal imaging

Optical coherence tomography angiography (OCTA) is a functional extension of optical coherence tomography for non-invasive in vivo three-dimensional imaging of the microvasculature of biological tissues. Several algorithms have been developed to construct OCTA images from the measured optical coherence tomography signals. In this study, we compared the performance of three OCTA algorithms that are based on the variance of phase, amplitude, and the complex representations of the optical coherence tomography signals for rodent retinal imaging, namely the phase variance, improved speckle contrast, and optical microangiography. The performance of the different algorithms was evaluated by comparing the quality of the OCTA images regarding how well the vasculature network can be resolved. Quantities that are widely used in ophthalmic studies including blood vessel density, vessel diameter index, vessel perimeter index, vessel complexity index were also compared. Results showed that both the improved speckle contrast and optical microangiography algorithms are more robust than phase variance, and they can reveal similar vasculature features while there are statistical differences in the calculated quantities.