This PDF file contains the front matter associated with SPIE Proceedings Volume 9541 including the Title Page, Copyright information, Table of Contents, Introduction, and Conference Committee listing.

Besides structural imaging, OCT can be used to estimate axial velocities of the sample resolved in depth by Doppler processing. In Fourier domain OCT (FD-OCT), this is accomplished by measuring the phase difference (i.e. phase shift) between timely separated A-scans at the same depth. In most cases, these data are disturbed by noise caused by intrinsic noise of the OCT system, specified by the SNR, and decorrelation noise caused by the transversal movement of the optical beam relative to the sample. Since the first use of Doppler methods in OCT, many methods to reduce the phase shift noise by averaging have been presented. While all these methods use a fixed set of consecutive A-scans, the best method, exhibiting no bias and having the smallest standard deviation, was questionable. Recently, Doppler processing methods depending on the mentioned noise sources and delivering the most likely phase shift and thereby axial velocity became available. The relation of these methods to previously known methods like the Kasai estimator, maximum likelihood estimator (MLE) and joint spectral and time domain OCT (jSTdOCT) will be discussed.

We describe a modification of a recently proposed unconventional OCT approach to 3D microvasculature imaging based on high-pass filtering of B-scans in the lateral direction. The B-scans are acquired in M-mode-like regime with highly overlapped A-scans. The goal of the described modification is to suppress non-fluid artifacts in the resultant microcirculation images. The modification is based on the amplitude normalization procedure of complex-valued OCT signal before subsequent processing. This allows one to efficiently suppress imaging degradation due to the influence of very bright spots/lines (e.g. from hairs on the surface) and retain images of real flows inside the tissue without any artificial cut-off of the surface signal, or application of pixel-intensity thresholds, or signal classification approaches.

This paper describes experiments and methodology for flow rate estimation using optical coherence tomography and dilution method in single fiber setup. The single fiber is created from custom made glass capillary and polypropylene hollow fiber. As a data source, measurements on single fiber phantom with continuous flow of carrier medium and bolus of Intralipid solution as a contrast agent were used using Thorlabs OCT OCS1300SS. The measured data were processed by methods of image processing, in order to precisely align the individual images in the sequence and extract dilution curves from the area inside the fiber. An experiment proved that optical coherence tomography can be used for flow rate estimation by the dilution method with precision around 7%.

One challenge that has long held the attention of scientists is that of clearly seeing objects hidden by turbid media, as smoke, fog or biological tissue, which has major implications in fields such as remote sensing or early diagnosis of diseases. Here, we combine structured incoherent illumination and bucket detection for imaging an absorbing object completely embedded in a scattering medium. A sequence of low-intensity microstructured light patterns is launched onto the object, whose image is accurately reconstructed through the light fluctuations measured by a single-pixel detector. Our technique is noninvasive, does not require coherent sources, raster scanning nor time-gated detection and benefits from the compressive sensing strategy. As a proof of concept, we experimentally retrieve the image of a transilluminated target both sandwiched between two holographic diffusers and embedded in a 6mm-thick sample of chicken breast.

By compensating the random phase delay acquired while a light beam crosses a scattering curtain, it is possible to address the light at selected target position beyond the obstacle. An opaque lens can produce foci with a resolution higher than conventional optics if a strongly scattering medium is exploited. In practice, subwavelength resolution is obtained only for weakly transmitting samples. Herein we present a method which allows obtaining tiny bright optical spots even in presence of a minimum amount of scattering (semi-transparent media) in the beam path. Using a High-Pass spatial filter we block the pseudo-ballistic components of the transmitted beam, we are able to gather light on a spot with a diameter which is one third of the typical speckle grain in absence of the filter.

We present a new concept for performing stimulated Raman spectroscopy and microscopy by employing rapidly wavelength swept Fourier Domain Mode locked (FDML) lasers [1]. FDML lasers are known for fastest imaging in swept-source optical coherence tomography [2, 3]. We employ this continuous and repetitive wavelength sweep to generate broadband, high resolution stimulated Raman spectra with a new, time-encoded (TICO) concept [4]. This allows for encoding and detecting the stimulated Raman gain on the FDML laser intensity directly in time. Therefore we use actively modulated pump lasers, which are electronically synchronized to the FDML laser, in combination with a fast analog-to-digital converter (ADC) at 1.8 GSamples/s. We present hyperspectral Raman images with color-coded, molecular contrast.

Optical-coherence tomography (OCT) is a promising non-invasive, high-resolution imaging modality which can be used for cancer diagnosis and its therapeutic assessment. However, speckle noise makes detection of cancer boundaries and image segmentation problematic and unreliable. Therefore, to improve the image analysis for a precise cancer border detection, the performance of different image processing algorithms such as mean, median, hybrid median filter and rotational kernel transformation (RKT) for this task is investigated. This is done on OCT images acquired from an ex-vivo human cancerous mucosa and in vitro by using cultivated tumour applied on organotypical hippocampal slice cultures. The preliminary results confirm that the border between the healthy and the cancer lesions can be identified precisely. The obtained results are verified with fluorescence microscopy. This research can improve cancer diagnosis and the detection of borders between healthy and cancerous tissue. Thus, it could also reduce the number of biopsies required during screening endoscopy by providing better guidance to the physician.

In vivo imaging technologies such as optical coherence tomography (OCT) and rodent models enable longitudinal studies of different ophthalmic pathologies and their underlying physiological changes. In this work a custom-made high resolution polarization sensitive OCT system is used to image the posterior eye of small rodents like mice and rats. Enface projections at certain depths/levels in the three-dimensional (3D) reflectivity images enable to visualize different vascular structures in the retina as well as in the choroid. Further, phase resolved motion contrast based OCT angiography allows to create vascular maps with a high contrast to distinguish between static tissue and dynamic scatterers such as red blood cells. The levels of the vascular maps are determined by a semi-automatic approach in which layers of the retina are segmented in a first step. In a second step the projection level can be adjusted by the operator to restrict the volume in which the en-face projection is carried out. To demonstrate the functionality of this approach rats and mice of different strains were imaged. The anesthetized rodents were scanned with an angle of up to 28° × 28°. Respiration induced motion artefacts were compensated by post-processing. The vascular maps based on the OCT reflectivity information as well as on the phase variance showed that the resolution of the custom-made systems is suitable to resolve small retinal capillaries in the outer plexiform layer. Further, larger vessels within the nerve fiber layer and the inner nuclear layer as well as in the choroid were visualized by the proposed approach.

Animal models play an important role for understanding the pathophysiology of glaucoma and age-related macular degeneration. With these models, longitudinal studies can be performed and therefore there is need for non-invasive evaluation of disease progress. For that purpose optical coherence tomography (OCT) can be used. Since tissues with different polarization properties are important in these diseases, polarization sensitive OCT (PS-OCT) could be a valuable tool in preclinical research. In this work a high resolution PS-OCT (HR-PS-OCT) system was used in-vivo for rodent retinal imaging. A super luminescent diode with a bandwidth of 100 nm was used as a light source that yielded an axial resolution of 5.1 μm in air (3.8 μm in tissue). The A-scan rate was 83 kHz, a whole 3D dataset was acquired in a few seconds (1536x1024x200 pixels in 3.5 s) which reduced motion artifacts. Rats (Sprague-Dawley, Long-Evans and Brown Norway) as well as mice (C57BL/6) were imaged. High resolution reflectivity images showed all retinal layers in all animals. From acquired data also phase retardation, fast axis orientation and degree of polarization uniformity (DOPU) images were calculated. On phase retardation images sclera was identified as birefringent and retinal pigment epithelium (RPE) and choroid as depolarizing tissues. Our results demonstrate the suitability of the system for high speed/resolution imaging in follow up studies on rodents.

Retinal laser photocoagulation represents a widely used treatment for retinal pathologies such as diabetic chorioretinopathy or diabetic edema. For effective treatment, an appropriate choice of the treatment energy dose is crucial to prevent excessive tissue damage caused by over-irradiation of the retina. In this manuscript we investigate simultaneous and time-resolved optical coherence tomography for its applicability to provide feedback to the ophthalmologist about the introduced retinal damage during laser photocoagulation. Time-resolved and volumetric optical coherence tomography data of 96 lesions on ex-vivo porcine samples, set with a 577 nm laser prototype and irradiance of between 300 and 8800 W=cm² were analyzed. Time-resolved scans were compared to volumetric scans of the lesion and correlated with ophthalmoscopic visibility. Lastly, image parameters extracted from optical coherence tomography Mscans, suitable for lesion classification were identified. Results presented in this work support the hypothesis that simultaneous optical coherence tomography provides valuable information about the extent of retinal tissue damage and may be used to guide retinal laser photocoagulation in the future.

Recently, optical coherence tomography (OCT) was utilized in multiple studies for structural and functional imaging of the middle ear and the tympanic membrane. Since Doppler OCT allows both, the spatially resolved measurement of the tympanic membrane oscillation and high-resolution imaging, it is regarded as a promising tool for future in vivo applications. In this study, Doppler OCT is utilized for the visualization of the tympanic membrane oscillation in temporal bones with simulated Eustachian catarrh, which was realized by generating a depression in the tympanic cavity. The transfer function, meaning the oscillation amplitude normalized to the applied sound pressure, is measured frequency resolved in the range from 0.5 kHz to 6 kHz and with a lateral spatial resolution of 0.4 mm. Typical oscillation patterns could be observed in case of ambient pressure in the tympanic cavity. Under depression the characteristic oscillation patterns were observed with widely congruent appearance but at higher frequencies.

The heart and its mechanical components, especially the heart valves and leaflets, are under enormous strain during lifetime. Like all highly stressed materials, also these biological components undergo fatigue and signs of wear, which impinge upon cardiac output and in the end on health and living comfort of affected patients. Thereby pathophysiological changes of the aortic valve leading to calcific aortic valve stenosis (AVS) as most frequent heart valve disease in humans are of particular interest. The knowledge about changes of the dynamic behavior during the course of this disease and the possibility of early stage diagnosis could lead to the development of new treatment strategies and drug-based options of prevention or therapy. ApoE-/- mice as established model of AVS versus wildtype mice were introduced in an ex vivo artificially stimulated heart model. 4D optical coherence tomography (OCT) in combination with high-speed video microscopy were applied to characterize dynamic behavior of the murine aortic valve and to characterize dynamic properties during artificial stimulation. OCT and high-speed video microscopy with high spatial and temporal resolution represent promising tools for the investigation of dynamic behavior and their changes in calcific aortic stenosis disease models in mice.

We present full volumetric high speed OCT imaging of the retina with multiple settings varying in volume size and volume rate. The volume size ranges from 255x255 A-scans to 160x40 A-scans with 450 samples per depth scan with volume rates varying between 20.8 V/s for the largest volumes to 195.2 V/s for the smallest. The system is based on a 1060nm Fourier domain mode locked (FDML) laser with 1.6MHz line rate. Scanning along the fast axis is performed with a 2.7 kHz or 4.3 kHz resonant scanner operated in bidirectional scanning mode, while a standard galvo scanner is used for the slow axis. The performance is analyzed with respect to various potential applications, like intraoperative OCT.

We tested and compared the capability of multiple optical coherence tomography (OCT) angiography methods: phase variance, amplitude decorrelation and speckle variance, with application of the split spectrum technique, to image the choroiretinal complex of the human eye. To test the possibility of OCT imaging stability improvement we utilized a real-time tracking scanning laser ophthalmoscopy (TSLO) system combined with a swept source OCT setup. In addition, we implemented a post- processing volume averaging method for improved angiographic image quality and reduction of motion artifacts. The OCT system operated at the central wavelength of 1040nm to enable sufficient depth penetration into the choroid. Imaging was performed in the eyes of healthy volunteers and patients diagnosed with age-related macular degeneration.

While FDML lasers with MHz sweep speeds have been presented five years ago, these devices have required manual control for startup and operation. Here, we present a fully self-starting and continuously regulated FDML laser with a sweep rate of 1.5 MHz. The laser operates over a sweep range of 115 nm centered at 1315 nm, and provides very high average output power of more than 100 mW. We characterize the laser performance, roll-off, coherence length and investigate the wavelength and phase stability of the laser output under changing environmental conditions. The high output power allows optical coherence tomography (OCT) imaging with an OCT sensitivity of 108 dB at 1.5 MHz.

Degradation of human ascending thoracic aorta has been visualized with Optical Coherence Tomography (OCT). OCT images of the vessel wall exhibit structural degradation in the media layer of the artery, being this disorder the final trigger of the pathology. The degeneration in the vessel wall appears as low-reflectivity areas due to different optical properties of acidic polysaccharides and mucopolysaccharides in contrast with typical ordered structure of smooth muscle cells, elastin and collagen fibers. An OCT dimension indicator of wall degradation can be generated upon the spatial quantification of the extension of degraded areas in a similar way as conventional histopathology. This proposed OCT marker can offer in the future a real-time clinical perception of the vessel status to help cardiovascular surgeons in vessel repair interventions. However, the delineation of degraded areas on the B-scan image from OCT is sometimes difficult due to presence of speckle noise, variable signal to noise ratio (SNR) conditions on the measurement process, etc. Degraded areas can be delimited by basic thresholding techniques taking advantage of disorders evidences in B-scan images, but this delineation is not optimum in the aorta samples and requires complex additional processing stages. This work proposes an optimized delineation of degraded areas within the aorta wall, robust to noisy environments, based on the iterative application of Otsu’s thresholding method. Results improve the delineation of wall anomalies compared with the simple application of the algorithm. Achievements could be also transferred to other clinical scenarios: carotid arteries, aorto-iliac or ilio-femoral sections, intracranial, etc.

The short-term jitter and longer-term wander of the frequency sweep profile of a Vernier-Tuned Distributed Bragg Reflector (VT-DBR) laser at 1550 nm used in OCT applications is characterized in this work. The VT-DBR has demonstrated success in source-swept OCT (SSOCT), performing both intensity [1] and phase-sensitive [2] OCT. The purpose of this paper is to investigate one of the unique aspects of the VT-DBR laser that makes it successful in OCT: the stability of the linear optical frequency sweep of the source. Jitter measurements of the optical frequency sweep are recorded using a 3-cavity 100 GHz free spectral range (FSR) solid etalon. A gas absorption reference cell is used for wander characterization. We report that the VT-DBR jitters by no more than 82 MHz RMS in optical frequency while sweeping at an 8 kHz repetition rate. Longer-term wander provides insight into the accuracy of the VT-DBR selfcalibration routine which produces an intrinsically linear optical frequency sweep. Over an 8-hour data collection period, the system maintains a linear sweep with an optical frequency step of 105 MHz per 2.5 ns with +/- 3 kHz per 2.5 ns (+/- 0.03%) peak-to-peak deviation. We find that the absolute frequency drifts by 325 MHz (2.6pm) over the same 8- hour period with ambient temperature fluctuations of no more than 5 °C. Results show that using calibration with a gas reference cell, picometer absolute wavelength accuracy of the laser can be achieved at any time for a single sweep. Stability and accuracy limits are thought to be due to electronic drive circuitry in the current design.

Non-contact imaging methods to distinguish between healthy tissue and brain tumor tissue during surgery would be highly desirable but are not yet available. Optical Coherence Tomography (OCT) is a non-invasive imaging technology with a resolution around 1-15 μm and a penetration depth of 1-2 mm that may satisfy the demands. To analyze its potential, we measured ex vivo human brain tumor tissue samples from 10 patients with a Spectral Domain OCT system (Thorlabs Callisto: center wavelength of 930 nm) and compared the results with standard histology. In detail, three different measurements were made for each sample. First the sample was measured directly after surgery. Then it was embedded in paraffin (also H and E staining) and examined for the second time. At last, the slices of each paraffin block cut by the pathology were measured. Each time a B-scan was created and for a better comparison with the histology a 3D image was generated, in order to get the corresponding en face images. In both, histopathological diagnosis and the analysis of the OCT images, different types of brain tumor showed difference in structure. This has been affirmed by two blinded investigators. Nevertheless the difference between two images of samples taken directly after surgery is less distinct. To enhance the contrast in the images further, we employ Spectroscopic OCT and pattern recognition algorithms and compare these results to the histopathological standard.

An algorithm for the simulation of Fourier domain optical coherence tomography (OCT) images of a cylinder based on an analytical solution of Maxwell's equations is presented. The characteristics of the simulated OCT signal are discussed and the whispering gallery modes as well as the geometrical optics signals from the cylinder are identified. An OCT scanner with an incident Gaussian beam is implemented to simulate two-dimensional B-scans.

When measuring static objects with 3D OCT, two opposing trends occur: If the integration time is too short, the measurement is noisy resulting in granulated textures on measured objects. If the integration time is too long, drifts e.g. due to thermal effects or unstable laser sources lead to blurred images. The Allan variance is a scheme to find the optimal integration time in terms of reducing noise without picking up signal drift. A long-term measurement with short integration time of a reference target under realistic conditions is needed to obtain the database for the calculation of the Allan variance. Longer integration times are simulated by taking averages of subsequent samples. The Allan variance being the mean of the squared differences between two consecutive averages is calculated for different integration times. The optimal integration time is achieved for minimal Allan variance. First, the scheme is explained and discussed with simulated data. Then, reference measurements of layers of adhesive tape made with a 3D OCT device are analysed to find the optimal integration time of the device. Finally, the findings are applied to the detection of water inclusions in calcite. With too short integration time the water inclusions appear with a stained surface. With the integration time increased towards the optimal time, the surfaces of the water inclusions get smoother and easier to discriminate from the background. Ready-to-use Octave code for the computation of the Allan variance is provided.

A general simulation framework has been developed for characterising eye motion artefacts in the ophthalmic imaging of ocular media. The optimisation of the scanning strategy for an optical coherence tomography of the anterior eye chamber has been presented as the first example of an application for the simulator.

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Optical Coherence Tomography (OCT) has shown a great potential as a complementary imaging tool in the diagnosis of skin diseases. Speckle noise is the most prominent artifact present in OCT images and could limit the interpretation and detection capabilities. In this work we propose a new speckle reduction process and compare it with various denoising filters with high edge-preserving potential, using several sets of dermatological OCT B-scans. To validate the performance we used a custom-designed spectral domain OCT and two different data set groups. The first group consisted in five datasets of a single B-scan captured N times (with N<20), the second were five 3D volumes of 25 Bscans. As quality metrics we used signal to noise (SNR), contrast to noise (CNR) and equivalent number of looks (ENL) ratios. Our results show that a process based on a combination of a 2D enhanced sigma digital filter and a wavelet compounding method achieves the best results in terms of the improvement of the quality metrics. In the first group of individual B-scans we achieved improvements in SNR, CNR and ENL of 16.87 dB, 2.19 and 328 respectively; for the 3D volume datasets the improvements were 15.65 dB, 3.44 and 1148. Our results suggest that the proposed enhancement process may significantly reduce speckle, increasing SNR, CNR and ENL and reducing the number of extra acquisitions of the same frame.