The trend towards homecare and point-of-care devices in medical care triggers the need for more compact and economic medical diagnostic technology. With this goal, we developed a flexible low-cost long-range SS-OCT capable of imaging the human eye from anterior to posterior segment, based on a single-mode tunable VCSEL at 850 nm. The system runs at A-scan rates of 2-25kHz with a measured maximum sensitivity of 97dB. In this work, we present in-vivo results of full eye imaging with an imaging range of 5 cm including both anterior and posterior segment B-scans.
A low-cost swept-source OCT system for retinal imaging was achieved, based on a thermally tuned vertical-cavity surface-emitting laser (VCSEL). Its center wavelength can be tuned by adjusting the operating temperature through modulation of the injection current. Sweep rates of 50-100 kHz with a sensitivity 97dB and an axial resolution of about 50μm in air have been achieved. We present the results of a human retina in vivo, using such a thermally tuned VCSEL-based SS-OCT system. Based on our results, we believe that this technology can be used as a cost-effective OCT alternative for point-of-care diagnostics.
The miniaturization of optical coherence tomography (OCT) systems could open up potential new markets, such as point-of-care application, home OCT to regularly monitor disease and treatment progress, and in low-resource settings. Photonic integrated circuits (PIC) are considered an attractive approach to miniaturize OCT. We present our recent achievements in in vivo retinal imaging with a PIC-based Mach-Zehnder interferometer integrated in a state-of-the-art ophthalmic OCT system. The system achieves 94 dB at 750 µW on the sample, running at 50 kHz. Preliminary results of a fully packaged 4-channel opto-electronic OCT engine further demonstrate the potential of PIC-based OCT.
A conditional generative adversarial network (cGAN) was developed to recover the depth resolution of optical coherence tomography images generated by a discontinuous spectrum in the visible wavelength range. Our approach was demonstrated on two phantoms and ex-vivo mouse ear tissue.
A novel non-iterative digital adaptive optics (DAO) technique is presented in which the wavefront error is calculated using the phase difference between the pupil field and its digital copies translated by a pixel along the horizontal and vertical direction of the defined co-ordinate of the pupil plane. Using this method, a large wavefront error with peak to valley of 20 radians and RMS error of 0.71 waves is detected and corrected in case of a micro-beads phantom sample. Also, human photoreceptor image is recovered from an aberrated retinal OCT acquired at the eccentricity of 2 degrees from fovea in-vivo with a flying spot OCT system using a phase stable akinetic swept source at 1060 nm.
In this pilot study, a multi-modal optical coherence microscopy (OCM) and fluorescence imaging system in the visible wavelength region was utilized to investigate the intra-tumor heterogeneity of lower grade brain tumor samples. OCM intensity data derived attenuation coefficients were calculated and cell density maps were created from intensity en-face projections. Sequentially preoperatively administrated 5-ALA fluorescence intensities were evaluated using the multi-modal setup. Optical imaging data results were compared to histology-derived cell density and cell proliferation rate maps.
Significance: Amyloid-beta (A-β) plaques are pathological protein deposits formed in the brain of Alzheimer’s disease (AD) patients upon disease progression. Further research is needed to elucidate the complex underlying mechanisms involved in their formation using label-free, tissue preserving, and volumetric techniques.
Aim: The aim is to achieve a one-to-one correlation of optical coherence tomography (OCT) data to histological micrographs of brain tissue using 1060-nm swept source OCT.
Approach: A-β plaques were investigated in ex-vivo AD brain tissue using OCT with the capability of switching between two magnifications. For the exact correlation to histology, a 3D-printed tool was designed to generate samples with parallel flat surfaces. Large field-of-view (FoV) and sequentially high-resolution volumes at different locations were acquired. The large FoV served to align the OCT to histology images; the high-resolution images were used to visualize fine details.
Results: The instrument and the presented method enabled an accurate correlation of histological micrographs with OCT data. A-β plaques were identified as hyperscattering features in both FoV OCT modalities. The plaques identified in volumetric OCT data were in good agreement with immunohistochemically derived micrographs.
Conclusion: OCT combined with the 3D-printed tool is a promising approach for label-free, nondestructive, volumetric, and fast tissue analysis.
Tunable laser sources with sweep-rates higher than 1MHz recently became commercially available. Today’s commercial ophthalmic OCT systems use sweep-rates in the 100-200kHz regime. These much faster laser sources can be used to either significantly reduce the imaging time or significantly increase the field of view (FOV). In this study we investigate the clinical value of OCT with MHz-rate swept source lasers. We implemented a versatile ophthalmic OCT system using a Frequency-Domain-Mode-Locked (FDML) laser with a sweep-rate of 1.7MHz, to address a variety of ophthalmic OCT imaging applications, exhibiting large imaging depth for wide field retinal OCT and OCT angiography (OCTA) with a field of view of up to 90 degrees, as well as for anterior segment imaging, and microscopic OCTA of the choriocapillaris with repetition rates of more than 1kHz.
We demonstrate imaging of amyloid-beta plaques in ex-vivo Alzheimer’s disease brain tissue using a 1060 nm swept source optical coherence tomography setup. This instrument enabled a global investigation of the brain tissue with a large field of view of 8 mm x 8 mm as well as sequential high resolution imaging. Amyloid-beta (A-β) plaques were identified as highly scattering features. Results were in good agreement with immunohistochemically stained images gained by histology.
A novel lipid sensitive OCT setup is presented, using light source with a central wavelength of 1280 nm and a spectral tuning range of 201 nm. A cholesterol plaque located in the aorta was imaged post mortem. Different spectral bands were chosen in post processing. Analyzing the signal attenuation of the different spectral bands enables us to see a clear difference between the lipid absorption in the plaque between the bands. In the normal tissue no clear separation of the signal attenuation can be found. This enables us to visualize the plaque on a three dimensional level.
We utilized a visible light optical coherence microscopy imaging (OCM) system to investigate the intra-tumor heterogeneity of glioblastoma samples. Using a broad visible spectrum an axial resolution of 0.8 μm was achieved. Objective lenses with short focal lengths were integrated to achieve high transverse resolution. Large field of view images were acquired using an automatic moving x-y-stage. Attenuation maps were calculated from OCM data and cell-density evaluations were performed from corresponding histological sections. Cell density and attenuation data were evaluated. Our analysis revealed intra-tumor heterogeneity by varying cell densities and attenuation coefficients throughout the tumorous areas.
The choriocappilaris layer is considered to be one of the first retinal layers affected in age related macular degeneration and other retinal diseases [1]. Imaging this fine vascular layer below the retinal pigment epithelium (RPE) with optical coherence tomography angiography (OCTA) has been very challenging due to the high scattering and absorption of the RPE. In OCTA higher inter B-scan times increase the sensitivity for slow motion and hence improve the contrast of fine vessels. However, it was demonstrated that for the choriocappilaris the opposite is the case [2]. Novel swept source laser technology enables acquiring images at A-scan-rates of 1.7 million A-scans per second, which is approx. 17 times faster than the fastest commercially available OCT devices [3, 4]. OCTA images of the human macula with different inter B-scan times were acquired and compared. The same prototype system was also used to acquire single shot wide-field OCTA images with up to 60 degree field of view.
Adaptive optics optical coherence tomography (AO-OCT) provides depth resolved images of the retina with cellular resolution [1, 2]. So far, various cell types have been visualized with this technique including rod photoreceptors [3], retinal pigment epithelium cells [3, 4] or Ganglion cells [5, 6]. However, a translation of this technology into clinical settings remains challenging as AOOCT systems are quite bulky and complex to operate. In addition, the clinical benefit of AO-OCT imaging has not yet been demonstrated as especially elderly patients are difficult to image. This presentation gives an overview over the performance of AO-OCT technology in a clinical setting.
Artificial test eyes have been developed for a multimodal ophthalmic imaging platform. The test eyes can be used for alignment of different imaging modalities and for fluorescence channel performance testing. Different scattering and absorption characteristics can be realized in the artificial retina.
5-aminolevulinic acid positive brain tumor samples were investigated with a multimodal visible light optical coherence microscope (OCM) and fluorescence imaging (FI) setup. Tumor specific contrast was achieved by the FI part while the three-dimensional tissue morphology was assessed by the OCM part of the setup.
We present a multimodal visible light optical coherence microscopy (OCM) and fluorescence imaging (FI) setup. Specification and phantom measurements were performed to characterize the system. Two applications in neuroimaging were investigated. First, curcumin-stained brain slices of a mouse model of Alzheimer’s disease were examined. Amyloid-beta plaques were identified based on the fluorescence of curcumin, and coregistered morphological images of the brain tissue were provided by the OCM channel. Second, human brain tumor biopsies retrieved intraoperatively were imaged prior to conventional neuropathologic work-up. OCM revealed the three-dimensional structure of the brain parenchyma, and FI added the tumor tissue-specific contrast. Attenuation coefficients computed from the OCM data and the florescence intensity values were analyzed and showed a statistically significant difference for 5-aminolevulinic acid (5-ALA)-positive and -negative brain tissues. OCM findings correlated well with malignant hot spots within brain tumor biopsies upon histopathology. The combination of OCM and FI seems to be a promising optical imaging modality providing complementary contrast for applications in the field of neuroimaging.
We implemented a combined visible light optical coherence microscopy (OCM) and fluorescence imaging platform. A supercontinuum light source in combination with a variable filter box (NKT Photonics) provided a spectral range of 425-680 nm. The OCM setup consists of a Michelson interferometer and a custom made spectrometer. Specification measurements were performed and an axial resolution of 0.88 μm in brain tissue was achieved. The transversal resolution was dependent on the objective lenses and varied from 2 to 8 μm. To change from OCM to fluorescence imaging, two mirrors had to be simply flipped into the light beam in the setup. For acquisition of fluorescence images, a photon multiplier tube (PMT) was used to detect light which had passed through a matched combination of emission, dichroic and excitation filters. As a first proof of concept, a fluorescence phantom consisting of curcumin powder mixed with mounting medium was imaged. The OCM images showed the three-dimensional structure of this phantom and specific contrast was gained by fluorescence imaging. As a control case, mounting medium without curcumin powder was imaged and no fluorescence was observed. One hallmark of Alzheimer's disease (AD) is the development of extracellular amyloid-beta plaques in the brain. The three-dimensional structure of these plaques was investigated with micrometer scale resolution using the OCM system. Curcumin can be used to specifically label amyloid-beta deposits. Curcumin stained brain slices of an AD mouse model were imaged and a specific contrast was gained by the fluorescence.
We exploit the intrinsic phase stability of akinetic swept source optical coherence tomography to demonstrate digital defocus correction in-vivo at a center wavelength of 1060nm. The high speed of 500kHz enables digital adaptive optics (AO) correction across a field of view of 1.8x1.5deg, currently limited by the employed galvo scanners. The source operates in a previously presented dual resolution mode OCT system (wide field >40deg, AO >3deg) with hardware based adaptive optics. The latter allows to efficiently combine hardware and digital AO, and to further optimize the AO imaging results. We demonstrate the digitally assisted AO performance for both structural imaging as well as for OCT angiography imaging across the full retina down to the choriocapillaris.
Investigations on retinal vasculature and blood flow are of interest for understanding and diagnostics of numerous ocular diseases. Conventional OCT systems use various scan patterns like linear B-scans, circular scans around the optic nerve head, or raster scans for 3D data acquisition. However, for some studies it is preferable to have customized scan patterns that can, e.g., follow the trace of an arbitrary linear structure in the retina, such as a vessel. In this work, we present an OCT instrument with an integrated retinal tracker that allows repeated scans along an arbitrary trace, whereby ocular motions are corrected by the retinal tracker. The setup comprises an OCT system and a line scanning laser ophthalmoscope (LSLO). The OCT subsystem operates at a center wavelength of 860 nm, with a bandwidth of 60 nm and an A-scan rate of 70kHz. The LSLO system operates at 790 nm and at a frame rate of 60 Hz. This system was used for reflectivity and Doppler imaging along retinal vessels. In a first step, the vessel is manually marked on the LSLO image. Then, repeated scans along the vessel trace are performed (2048 A-scans per scan along trace, up to 500 scans along the trace). The intensity images show a clear delineation of the vessel walls, the phase difference (Doppler) tomograms allow for a time-resolved analysis of blood flow along the vessel over the cardiac cycle.
KEYWORDS: Optical coherence tomography, Endoscopy, Endoscopes, Signal detection, In vivo imaging, Mirrors, GRIN lenses, Tissues, Image quality, Imaging systems
We demonstrate in vivo endoscopic optical coherence tomography (OCT) imaging in the forward direction using a flexible fiber bundle (FB). In comparison to current conventional forward-looking probe schemes, our approach simplifies the endoscope design by avoiding the integration of any beam steering components in the distal probe end due to two-dimensional scanning of a focused light beam over the proximal FB surface. We describe the challenges that arise when OCT imaging with an FB is performed, such as multimoding or cross coupling. The performance of different FBs varying in parameters, such as numerical aperture, core size, core structure, and flexibility, was consequently compared, and image quality degrading artifacts were described in detail. Based on our findings, we propose an optimal FB design for endoscopic OCT imaging.
Adaptive Optics (AO) retinal imaging is revealing microscopic structures of the eye in a non-invasive way. Due to anisoplanatism, conventional AO systems are efficient on small 1°x1° field of view (FoV). We present a lens-based AO scanning laser ophthalmoscope (SLO) set-up with 2 deformable mirrors (DM), providing high-resolution retinal imaging on a 4°x4° FoV, for an eye pupil diameter of 7 mm. The first DM is in a pupil plane and is driven using a Shack-Hartmann (SH). The second DM is conjugated to a plane located 0.7 mm in front of the retina, to correct for aberrations varying within the FoV. Its shape is optimized using sensorless AO technique.
The performance of this set-up was characterized in-vivo by measuring the eyes of four healthy volunteers. The obtained image quality was satisfactory and uniform over the entire FoV. Foveal cones could be resolved and no image distortion was detected. Furthermore, a 10°x10° FoV image was acquired at the fovea of one volunteer, by stitching 9 images recorded at different eccentricities. Finally, different layers of the retina were imaged. In addition to the photoreceptors mosaic, small capillaries and nerve fibers were clearly identified.
The presented AO-SLO instrument provides high-resolution images of the retina on a relatively large FoV in reasonable time. With 2 DMs, one SH and no guide star, the system stays quite simple. The imaging performance of the set-up was validated on 4 healthy volunteers and we are currently imaging patients with different eye diseases.
In this work we investigate the benefits of using optical coherence tomography angiography (OCTA) in combination with adaptive optics (AO) technology. It has been demonstrated that the contrast of vessels and small capillaries can be greatly enhanced by the use of OCTA. Moreover, small capillaries that are below the transverse resolution of the ophthalmic instrument can be detected. This opens unique opportunities for diagnosing retinal diseases. However, there are some limitations of this technology such as shadowing artifacts caused by overlying vasculature or the inability to determine the true extension of a vessel. Thus, the evaluation of the vascular structure and density can be misleading. To overcome these limitations we applied the OCT angiography technique to images recorded with AO-OCT. Due to the higher collection efficiency of AO-OCT in comparison with standard OCT an increased intensity contrast of vasculature can be seen. Using AO-OCTA the contrast of the vasculature to the surrounding static tissue is further increased. The improved transverse resolution and the reduced depth of focus of the AO-OCT greatly reduce shadowing artifacts allowing for a correct differentiation and segmentation of different vascular layers of the inner retina. The method is investigated in healthy volunteers and in patients with diabetic retinopathy.
We present the optimization of an adaptive optics loop for retinal imaging. Generally, the wave-front is overdetermined compared to the number of corrector elements. The sampling of the sensor can be reduced while maintaining an efficient correction, leading to higher sensitivity, faster correction and larger dynamic range. An analytical model was developed to characterize the link between number of actuators, number of micro-lenses and correction performance. The optimized correction loop was introduced into a scanning laser ophthalmoscope. In vivo images of foveal photoreceptors were recorded and the obtained image quality is equivalent to the state of the art in retinal AO-imaging.
Adaptive optics (AO) is essential in order to visualize small structures such as cone and rod photoreceptors in the living human retina in vivo. By combining AO with optical coherence tomography (OCT) the axial resolution in the images can be further improved. OCT provides access to the phase of the light returning from the retina which allows a measurement of subtle length changes in the nanometer range. These occur for example during the renewal process of cone outer segments. We present an approach for measuring very small length changes using an extended AO scanning laser ophthalmoscope (SLO)/ OCT instrument. By adding a second OCT interferometer that shares the same sample arm as the first interferometer, phase sensitive measurements can be performed in the en-face imaging plane. Frame averaging decreases phase noise which greatly improves the precision in the measurement of associated length changes.
Multi-object adaptive optics requires a tomographic reconstructor to compute the AO correction for scientific targets
within the field, using measurements of incoming turbulence from guide stars angularly separated from the science
targets. We have developed a reconstructor using an artificial neural network, which is trained in simulation only.
We obtained similar or better results than current reconstructors, such as least-squares and Learn and Apply, in
simulation and also tested the new technique in the laboratory. The method is robust and can cope well with
variations in the atmospheric conditions. We present the technique, our latest results and plans for a full MOAO experiment.
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