This study tested the utility of optical coherence tomography (OCT)-based indentation to assess mechanical properties of respiratory tissues in disease. Using OCT-based indentation, the elastic modulus of mouse diaphragm was measured from changes in diaphragm thickness in response to an applied force provided by an indenter. We used a transgenic mouse model of chronic lung disease induced by the overexpression of transforming growth factor-alpha (TGF-±), established by the presence of pleural and peribronchial fibrosis and impaired lung mechanics determined by the forced oscillation technique and plethysmography. Diaphragm elastic modulus assessed by OCT-based indentation was reduced by TGF-± at both left and right lateral locations (p
Imaging of alveoli in situ has for the most part been infeasible due to the high resolution required to discern individual alveoli and limited access to alveoli beneath the lung surface. In this study, we present a novel technique to image alveoli using optical coherence tomography (OCT). We propose the use of OCT needle probes, where the distal imaging probe has been miniaturized and encased within a hypodermic needle (as small as 30-gauge, outer diameter 310 Î¼m), allowing insertion deep within the lung tissue with minimal tissue distortion. Such probes enable imaging at a resolution of
Angular diversity is a successful speckle-reduction technique in optical coherence tomography (OCT). We employ angle-dependent detection for a different purpose: to distinguish the singly backscattered and multiply scattered signal components. Single backscattering is highly correlated over a large range of detection angles; multiple scattering rapidly decorrelates as the angle is varied. Theoretical justification is provided using a linear-systems description of the OCT imaging process; detection of multiple scattering is corroborated experimentally. Â
Probing the mechanical properties of tissue on the microscale could aid in the identification of diseased tissues that are inadequately detected using palpation or current clinical imaging modalities, with potential to guide medical procedures such as the excision of breast tumours. Compression optical coherence elastography (OCE) maps tissue strain with microscale spatial resolution and can delineate microstructural features within breast tissues. However, without a measure of the locally applied stress, strain provides only a qualitative indication of mechanical properties. To overcome this limitation, we present quantitative micro-elastography, which combines compression OCE with a compliant stress sensor to image tissue elasticity. The sensor consists of a layer of translucent silicone with well-characterized stress-strain behaviour. The measured strain in the sensor is used to estimate the two-dimensional stress distribution applied to the sample surface. Elasticity is determined by dividing the stress by the strain in the sample. We show that quantification of elasticity can improve the ability of compression OCE to distinguish between tissues, thereby extending the potential for inter-sample comparison and longitudinal studies of tissue elasticity. We validate the technique using tissue-mimicking phantoms and demonstrate the ability to map elasticity of freshly excised malignant and benign human breast tissues.
To the best of our knowledge, we present the first needle probe for combined optical coherence tomography (OCT), and fluorescence imaging. The probe uses double-clad fiber (DCF) that guides the OCT signal and fluorescence excitation light in the core and collects and guides the returning fluorescence in the large-diameter multimode inner cladding. It is interfaced to a 1310 nm swept-source OCT system that has been modified to enable simultaneous 488 nm fluorescence excitation and >500 nm emission detection by using a DCF coupler to extract the returning fluorescence signal in the inner cladding with high efficiency. We present imaging results from an excised sheep lung with fluorescein solution infused through the vasculature. We were able to identify alveoli, bronchioles, and blood vessels. The results demonstrate that the combined OCT plus fluorescence needle images provide improved tissue differentiation over OCT alone.
We present a high-optical-quality imaging needle for optical coherence tomography (OCT) that achieves sensitivity and resolution comparable to conventional free-space OCT sample arms. The side-viewing needle design utilizes total internal reflection from an angle-polished fiber tip, encased in a glass microcapillary. Fusion of the capillary to the fiber provides a robust, optical-quality output window. The needle's focusing optics are based on an astigmatism-free design, which exploits the "focal shift" phenomenon for focused Gaussian beams to achieve equal working distances (WDs) for both axes. We present a fabricated needle with a WD ratio of 0.98 for imaging in an aqueous environment. Our needle achieves the highest sensitivity of currently reported OCT imaging needles (112 dB), and we demonstrate its performance by superficial imaging of human skin and 3D volumetric imaging within a biological sample.
We present an acquisition method for optical coherence elastography (OCE) that enables acquisition of three-dimensional elastograms in 5 s, an order of magnitude faster than previously reported. In this method, based on compression elastography, the mechanical load applied to the sample is altered between acquisitions of consecutive optical coherence tomography volume scans (C-scans). The voxel-by-voxel phase difference between the volumes is used to determine the axial displacement and determining the gradient of the axial displacement versus depth gives the local axial strain. We demonstrate sub-100-microstrain sensitivity and high contrast in elastograms, acquired in 5 s, of structured phantoms and freshly excised rat muscle tissue that are comparable in strain sensitivity and dynamic range to our previously reported B-scan-based method. The much higher acquisition speed may expedite the translation of OCE to clinical and in vivo applications
Endoscopic treatment of lower airway pathologies requires accurate quantification of airway dimensions. We demonstrate the application of a real-time endoscopic optical coherence tomography system that can image lower airway anatomy and quantify airway lumen dimensions intra-operatively. Results demonstrate the ability to acquire 3D scans of airway anatomy and include comparison against a pre-operative X-ray CT. The paper also illustrates the capability of the system to assess the real-time dynamic changes within the airway that occur during respiration.
Es'haghian S., Gong P., Chin L., Harms K.-A., Murray A., Rea S., Kennedy B.F., Wood F.M., Sampson David, McLaughlin R.A. (2017) Investigation of optical attenuation imaging using optical coherence tomography for monitoring of scars undergoing fractional laser treatment, Journal of Biophotonics 10 (4) pp. 511-522
We demonstrate the use of the near-infrared attenuation coefficient, measured using optical coherence tomography (OCT), in longitudinal assessment of hypertrophic burn scars undergoing fractional laser treatment. The measurement method incorporates blood vessel detection by speckle decorrelation and masking, and a robust regression estimator to produce 2D en face parametric images of the attenuation coefficient of the dermis. Through reliable co-location of the field of view across pre- and post-treatment imaging sessions, the study was able to quantify changes in the attenuation coefficient of the dermis over a period of ~20 weeks in seven patients. Minimal variation was observed in the mean attenuation coefficient of normal skin and control (untreated) mature scars, as expected. However, a significant decrease (13 ± 5%, mean ± standard deviation) was observed in the treated mature scars, resulting in a greater distinction from normal skin in response to localized damage from the laser treatment. By contrast, we observed an increase in the mean attenuation coefficient of treated (31 ± 27%) and control (27 ± 20%) immature scars, with numerical values incrementally approaching normal skin as the healing progressed. This pilot study supports conducting a more extensive investigation of OCT attenuation imaging for quantitative longitudinal monitoring of scars. (Figure presented.) En face 2D OCT attenuation coefficient map of a treated immature scar derived from the pre-treatment (top) and the post-treatment (bottom) scans. (Vasculature (black) is masked out.) The scale bars are 0.5 mm.
We have developed an anatomical optical coherence tomography system for imaging the human upper airway in vivo. We describe an example of clinical research currently underway with this system; the ability to measure the change in airway dimensions due to anatomical position. Although the system imaging range is well-matched to a typical airway, we have observed a range of conditions which preclude the capture of full airway profiles in a small number of cases. Here, we demonstrate an improvement in the system range which allows us to successfully measure a larger percentage of subjects.
A scheme for all-optical conversion of codes for coherent code-division multiplexing is proposed. The key functionality of the scheme is demonstrated by error-free conversion of an encoded data stream at 10 Gbit/s from one code to another. The scheme may enable routing and add/drop multiplexing based on optical codes.
We review the development of phantoms for optical coherence tomography (OCT) designed to replicate the optical, mechanical and structural properties of a range of tissues. Such phantoms are a key requirement for the continued development of OCT techniques and applications. We focus on phantoms based on silicone, fibrin and poly(vinyl alcohol) cryogels (PVA-C), as we believe these materials hold the most promise for durable and accurate replication of tissue properties.
Multiple scattering is one of the main degrading influences in optical coherence tomography, but to date its presence in an image can only be indirectly inferred. We present a polarization-sensitive method that shows the potential to detect it more directly, based on the degree to which the detected polarization state at any given image point is correlated with the mean state over the surrounding region. We report the validation of the method in microsphere suspensions, showing a strong dependence of the degree of correlation upon the extent to which multiply scattered light is coherently detected. We demonstrate the method's utility in various tissues, including chicken breast ex vivo and human skin and nailfold in vivo.
Three-dimensional optical coherence tomography (3D-OCT) is used to evaluate the structure and pathology of regenerating mouse skeletal muscle autografts for the first time. The death of myofibers with associated inflammation and subsequent new muscle formation in this graft model represents key features of necrosis and inflammation in the human disease Duchenne muscular dystrophy. We perform 3D-OCT imaging of excised autografts and compare OCT images with coregistered histology. The OCT images readily distinguish the necrotic and inflammatory tissue of the graft from the intact healthy muscle fibers in the underlying host tissue. These preliminary findings suggest that, with further development, 3D-OCT could be used as a tool for the evaluation of small-animal muscle morphology and pathology, in particular, for analysis of mouse models of muscular dystrophy.
Evaluation of lymph node involvement is an important factor in detecting metastasis and deciding whether to perform axillary lymph node dissection (ALND) in breast cancer surgery. As ALND is associated with potentially severe long term morbidity, the accuracy of lymph node assessment is imperative in avoiding unnecessary ALND. The mechanical properties of malignant lymph nodes are often distinct from those of normal nodes. A method to image the micro-scale mechanical properties of lymph nodes could, thus, provide diagnostic information to aid in the assessment of lymph node involvement in metastatic cancer. In this study, we scan axillary lymph nodes, freshly excised from breast cancer patients, with optical coherence micro-elastography (OCME), a method of imaging micro-scale mechanical strain, to assess its potential for the intraoperative assessment of lymph node involvement.
Twenty-six fresh, unstained lymph nodes were imaged from 15 patients undergoing mastectomy or breast-conserving surgery with axillary clearance. Lymph node specimens were bisected to allow imaging of the internal face of each node. Co-located OCME and optical coherence tomography (OCT) scans were taken of each sample, and the results compared to standard post-operative hematoxylin-and-eosin-stained histology.
The optical backscattering signal provided by OCT alone may not provide reliable differentiation by inspection between benign and malignant lymphoid tissue. Alternatively, OCME highlights local changes in tissue strain that correspond to malignancy and are distinct from strain patterns in benign lymphoid tissue. The mechanical contrast provided by OCME complements the optical contrast provided by OCT and aids in the differentiation of malignant tumor from uninvolved lymphoid tissue.
The combination of OCME and OCT images represents a promising method for the identification of malignant lymphoid tissue. This method shows potential to provide intraoperative assessment of lymph node involvement, thus, preventing unnecessary removal of uninvolved tissues and improving patient outcomes.
Purpose: To evaluate the impact of image magnification correction on superficial retinal vessel density (SRVD) and foveal avascular zone area (FAZA) measurements using optical coherence tomography angiography (OCTA).
Methods: Participants with healthy retinas were recruited for ocular biometry, refraction, and RTVue XR Avanti OCTA imaging with the 3 × 3-mm protocol. The foveal and parafoveal SRVD and FAZA were quantified with custom software before and after correction for magnification error using the Littman and the modified Bennett formulae. Relative changes between corrected and uncorrected SRVD and FAZA were calculated.
Results: Forty subjects were enrolled and the median (range) age of the participants was 30 (18?74) years. The mean (range) spherical equivalent refractive error was ?1.65 (?8.00 to +4.88) diopters and mean (range) axial length was 24.42 mm (21.27?28.85). Images from 13 eyes were excluded due to poor image quality leaving 67 for analysis. Relative changes in foveal and parafoveal SRVD and FAZA after correction ranged from ?20% to +10%, ?3% to +2%, and ?20% to +51%, respectively. Image size correction in measurements of foveal SRVD and FAZA was greater than 5% in 51% and 74% of eyes, respectively. In contrast, 100% of eyes had less than 5% correction in measurements of parafoveal SRVD.
Conclusions: Ocular biometry should be performed with OCTA to correct image magnification error induced by axial length variation. We advise caution when interpreting interocular and interindividual comparisons of SRVD and FAZA derived from OCTA without image size correction.
Needle-based devices, which are in wide clinical use for needle biopsy procedures, may be augmented by suitable optical techniques for the localization and diagnosis of diseased tissue. Tissue refractive index is one optical contrast mechanism with diagnostic potential. In the case of mammary tissue, for example, recent research indicates that refractive index variations between tissue types may be useful for the identification of cancerous tissue. While many coherence-based forward-sensing devices have been developed to detect scattering changes, none have demonstrated refractive index measurement capabilities. We present a novel needle-based device that is capable of simultaneously measuring refractive index and scattering. Coupled to the sample arm of an optical coherence tomography system, the needle device detects the scattering response and optical pathlength through tissue residing in a fixed-width channel. Near-infrared measurements of tissues and materials with known optical properties using a prototype device will be presented. This work demonstrates the feasibility of integrated in vivo measurement of refractive index and scattering in conjunction with existing clinical needle-based devices.
We utilize Fourier-holographic light scattering angular spectroscopy to record the spatially resolved complex angular scattering spectra of samples over wide fields of view in a single or few image captures. Without resolving individual scatterers, we are able to generate spatially-resolved particle size maps for samples composed of spherical scatterers, by comparing generated spectra with Mie-theory predictions. We present a theoretical discussion of the fundamental principles of our technique and, in addition to the sphere samples, apply it experimentally to a biological sample which comprises red blood cells. Our method could possibly represent an efficient alternative to the time-consuming and laborious conventional procedure in light microscopy of image tiling and inspection, for the characterization of microscopic morphology over wide fields of view.
We report single-measurement, full-range imaging of local polarization properties in the human anterior segment in vivo with polarization-sensitive optical coherence tomography (PS-OCT). Off-pivot galvanometer-mirror phase shifting used to extend the system's axial imaging range sufficiently to reconstruct local polarization properties of the anterior segment.
We investigate, both theoretically and experimentally, the signal-to-noise ratio (SNR) of modulated amplified spontaneous emission (ASE) transmitted over dispersive fiber. We observe two significant effects; firstly, the signal-to-excess-noise ratio (SNR ex) varies across the pulse reaching its maximum value near the peak of the detected signal; and secondly, this maximum value decreases with increasing fiber dispersion-induced pulse broadening. Accurate calculation of transmission performance of high bit-rate optical communication systems employing ASE sources, such as spectrum slicing, requires inclusion of these effects.
Hull thickness is an important component of seed quality, which effects dehulling ability, feed or food nutritional aspects and cooking times. A breeding objective in Lupinus angustifolius crop improvement is to reduce hull thickness and a rapid screening method is needed to efficiently screen genotypes. Optical coherence tomography (OCT) imaging using infrared illumination at 980 nm was used to compare hull thickness of genotypes of four lupin species. OCT-derived hull layer thickness correlated highly with actual hull thickness determined by environmental scanning electron microscopy (r = 0.90) and allowed reliable distinction between mutant (thin-hulled) and parent genotypes of L. angustifolius. The imaging could clearly penetrate lupin seed to a depth of approximately 200 ¼m. The use of OCT to measure hull thickness has the advantage that it is rapid and non-destructive and should be very useful in selecting thin hull lines of lupins and other species on a single seed basis in germplasm or progeny from crosses.
We report the effects on two-photon excitation microscopy of applying optical clearing agents to human skin tissue samples. We demonstrate that the agents glycerol, propylene glycol and glucose in aqueous solution are all effective in enhancing penetration depth (by up to a factor of 2) and in increasing image contrast (by up to a factor of 90 at 80 ¼m depth) in 150 ¼m thick sections. We analysed the dynamics of the clearing process, by developing a simple theoretical model based on the free diffusion of the agent into the tissue. In experiments employing simultaneous two-photon excitation and second harmonic generation microscopy similar contrast was produced. A preliminary measurement of the clearing effect on a bulk skin sample is also presented. All three agents are potentially biocompatible and effective in reducing scattering; hence, in improving light penetration depth and image contrast. As such, they could be suitable for in vivo application in two-photon microscopy, as well as in other techniques performing optical biopsy of human skin tissue.
We demonstrate the modification of optical coherence elastography to advance from relative strain images to quantified tissue stiffness on the micro-scale. We highlight the nonlinear dependence of tissue stiffness on the applied load and consider how nonlinearity may help characterise soft tissues.
This study presents the first in vivo longitudinal assessment of scar vasculature in ablative fractional laser treatment using optical coherence tomography (OCT). A method based on OCT speckle decorrelation was developed to visualize and quantify the scar vasculature over the treatment period. Through reliable co?location of the imaging field of view across multiple imaging sessions, and compensation for motion artifact, the study was able to track the same scar tissue over a period of several months, and quantify changes in the vasculature area density. The results show incidences of occlusion of individual vessels 3 days after the first treatment. The subsequent responses Ü20 weeks after the initial treatment show differences between immature and mature scars. Image analysis showed a distinct decrease (25 ± 13%, mean ± standard deviation) and increase (19 ± 5%) of vasculature area density for the immature and mature scars, respectively. This study establishes the feasibility of OCT imaging for quantitative longitudinal monitoring of vasculature in scar treatment.
We demonstrate a multiwavelength source of amplified spontaneous emission suitable for spectrum-sliced, wavelength-division multiplexed access and local-area networks. The source, based on flat-gain optical amplification of a Fabry-Perot laser biased below threshold, provides up to 30-wavelength channels spaced by 138 GHz. We show the potential of the source in network applications by demonstrating wavelength-stabilized, low (<10-9) error rate operation of all 30 wavelength channels at 622 Mb/s.
We report on the design and implementation of a gradient-index microendoscope suitable for accessing tissues deep within the body using confocal fluorescence imaging. The 350-¼m diameter microendoscope has a length of 27 mm, which enables it to be inserted through a 22-gauge hypodermic needle. A prototype imaging system is demonstrated to obtain images of tissue samples at depths of ~15 mm with a lateral resolution of ~700 nm. To the best of our knowledge, this is the highest resolution and imaging depth reported for a confocal probe of these dimensions. We employ a scanning arrangement using a lensed fiber that can conveniently control the input beam parameters without causing off-axis aberrations typically present in the optical relay lenses used in galvanometer-mirror scanning systems.
Fiber-optic needle probes are highly miniaturized imaging devices that enable imaging deep inside the body. Utilizing optical coherence tomography (OCT), these devices replace the standard scanning mechanism of an OCT scanner with all-fiber focusing optics small enough to be encased within a hypodermic needle. We describe recent innovation in the design of these probes, including novel fiber-optic configurations to achieve extended depth of focus, and the use of double-clad fiber to enable the first dual-modality fiber-optic needle probe, simultaneously acquiring both OCT and fluorescence images.
Optical coherence tomography is a rapidly maturing optical imaging technology, enabling study of the in vivo structure of lung tissue at a scale of tens of micrometers. It has been used to assess the layered structure of airway walls, quantify both airway lumen caliber and compliance, and image individual alveoli. This article provides an overview of the technology and reviews its capability to provide new insights into respiratory disease.
The transmission of a spectrum-sliced WDM channel at 622 Mbit/s over 60 km of nondispersion-shifted fibre using an optical bandwidth of only 0.23 nm is reported. This is the highest single channel bit rate-length product (40Gbit/s-km) and smallest channel bandwidth reported to date for spectrum-sliced WDM systems. The bit error rate performance is theoretically predicted and experimentally confirmed and limits on the bit rate-length products of spectrum-sliced WDM channels using nondispersion-shifted fibre in the 1550nm window are given.
The real-time dispersion compensation in scanning interferometry was discussed. The static grating tilt in a scanning frequency-domain optical delay line was shown to produce dispersion that was linearly proportional to scan position. It was found that this device has application in scanning interferometry in general and in optical coherence tomography (OCT) in particular.
Lucey A.D., King A.J.C., Tetlow G.A., Wang J., Armstrong J.J., Leigh M.S., Paduch A., Walsh J.H., Sampson D.D., Eastwood P.R., Hillman D.R. (2010) Measurement, reconstruction, and flow-field computation of the human pharynx with application to sleep apnea, IEEE Transactions on Biomedical Engineering 57 (10 PAR) pp. 2535-2548
Repetitive closure of the upper airway characterizes obstructive sleep apnea. It disrupts sleep causing excessive daytime drowsiness and is linked to hypertension and cardiovascular disease. Previous studies simulating the underlying fluid mechanics are based upon geometries, time-averaged over the respiratory cycle, obtained usually via MRI or CT scans. Here, we generate an anatomically correct geometry from data captured in vivo by an endoscopic optical technique. This allows quantitative real-time imaging of the internal cross section with minimal invasiveness. The steady inhalation flow field is computed using a k- shear-stress transport (SST) turbulence model. Simulations reveal flow mechanisms that produce low-pressure regions on the sidewalls of the pharynx and on the soft palate within the pharyngeal section of minimum area. Soft-palate displacement and side-wall deformations further reduce the pressures in these regions, thus creating forces that would tend to narrow the airway. These phenomena suggest a mechanism for airway closure in the lateral direction as clinically observed. Correlations between pressure and airway deformation indicate that quantitative prediction of the low-pressure regions for an individual are possible. The present predictions warrant and can guide clinical investigation to confirm the phenomenology and its quantification, while the overall approach represents an advancement toward patient-specific modeling.
We present optical coherence micro-elastography, an improved form of compression optical coherence elastography. We demonstrate the capacity of this technique to produce en face images, closely corresponding with histology, that reveal micro-scale mechanical contrast in human breast and lymph node tissues. We use phase-sensitive, three-dimensional optical coherence tomography (OCT) to probe the nanometer-to-micrometer-scale axial displacements in tissues induced by compressive loading. Optical coherence micro- elastography incorporates common-path interferometry, weighted averaging of the complex OCT signal and weighted least-squares regression. Using three-dimensional phase unwrapping, we have increased the maximum detectable strain eleven-fold over no unwrapping and the minimum detectable strain is 2.6 Î¼Îµ. We demonstrate the potential of mechanical over optical contrast for visualizing micro-scale tissue structures in human breast cancer pathology and lymph node morphology.
We report a new synthetic aperture optical microscopy in which high-resolution, wide-field amplitude and phase images are synthesized from a set of Fourier holograms. Each hologram records a region of the complex two-dimensional spatial frequency spectrum of an object, determined by the illumination field's spatial and spectral properties and the collection angle and solid angle. We demonstrate synthetic microscopic imaging in which spatial frequencies that are well outside the modulation transfer function of the collection optical system are recorded while maintaining the long working distance and wide field of view.
Polarization-sensitive OCT (PS-OCT) has proven useful in determining the stress-induced birefringence of non-biological materials, but such utility in biological tissues subjected to stress has not been well studied yet. To study stress-induced birefringence of biological tissues, we use a swept-source PS-OCT system with passively depth-encoded, orthogonal polarization states in the illumination path and polarization-diversity detection and a Mueller formalism in post-processing. We present measurements of stress-induced changes in the birefringence of non-biological and biological samples that provide useful benchmarks in further assessing the utility of this approach.
Optical elastography, the use of optics to characterize and map the mechanical properties of biological tissue, involves measuring the deformation of tissue in response to a load. Such measurements may be used to form an image of a mechanical property, often elastic modulus, with the resulting mechanical contrast complementary to the more familiar optical contrast. Optical elastography is experiencing new impetus in response to developments in the closely related fields of cell mechanics and medical imaging, aided by advances in photonics technology, and through probing the microscale between that of cells and whole tissues. Two techniques ? optical coherence elastography and Brillouin microscopy ? have recently shown particular promise for medical applications, such as in ophthalmology and oncology, and as new techniques in cell mechanics.
High-resolution tactile imaging, superior to the sense of touch, has potential for future biomedical applications such as robotic surgery. In this paper, we propose a tactile imaging method, termed computational optical palpation, based on measuring the change in thickness of a thin, compliant layer with optical coherence tomography and calculating tactile stress using finite-element analysis. We demonstrate our method on test targets and on freshly excised human breast fibroadenoma, demonstrating a resolution of up to 15?25 µm and a field of view of up to 7 mm. Our method is open source and readily adaptable to other imaging modalities, such as ultrasonography and confocal microscopy.
We show for the first time, to our knowledge, high-resolution wide-field images of biological samples recorded using coherent aperture-synthesis Fourier holography. To achieve this, we combined off-axis plane-wave polarized illumination with an axial sample rotation and polarization-sensitive collection of backscat- tered light. We synthesized 180 Fourier holograms using an efficient postdetection phase-matching correlation scheme. The result was an annular spatial frequency-space synthetic aperture (NA=0.93) with an effective area 25 times larger than that due to a single hologram. A high-resolution high-contrast microscopic reconstruction of biological tissue was computed over a sample area of 9mm2 from holograms acquired at 34 mm working distance.
We incorporate, for the first time, optical coherence elastography (OCE) into a needle probe and demonstrate its ability to measure the microscopic deformation of soft tissues located well beyond the depth limit of reports to date. Needle OCE utilizes the force imparted by the needle tip as the loading mechanism and measures tissue deformation ahead of the needle during insertion. Measurements were performed in tissue-mimicking phantoms and ex vivo porcine trachea. Results demonstrate differentiation of tissues based on mechanical properties and highlight the potential of needle OCE for in vivo tissue boundary detection.
Cellular-scale imaging of the mechanical properties of tissue has helped to reveal the origins of disease; however, cellular-scale resolution is not readily achievable in intact tissue volumes. Here, we demonstrate volumetric imaging of Young?s modulus using ultrahigh-resolution optical coherence elastography, and apply it to characterizing the stiffness of mouse aortas. We achieve isotropic resolution of better than 15 ¼m over a 1-mm lateral field of view through the entire depth of an intact aortic wall. We employ a method of quasi-static compression elastography that measures volumetric axial strain and uses a compliant, transparent layer to measure surface axial stress. This combination is used to estimate Young?s modulus throughout the volume. We demonstrate differentiation by stiffness of individual elastic lamellae and vascular smooth muscle. We observe stiffening of the aorta in regulator of G protein signaling 5-deficient mice, a model that is linked to vascular remodeling and fibrosis. We observe increased stiffness with proximity to the heart, as well as regions with micro-structural and micro-mechanical signatures characteristic of fibrous and lipid-rich tissue. High-resolution imaging of Young?s modulus with optical coherence elastography may become an important tool in vascular biology and in other fields concerned with understanding the role of mechanics within the complex three-dimensional architecture of tissue.
Optical coherence elastography employs optical coherence tomography (OCT) to measure the displacement of tissues under load and, thus, maps the resulting strain into an image, known as an elastogram. We present a new improved method to measure vibration amplitude in dynamic optical coherence elastography. The tissue vibration amplitude caused by sinusoidal loading is measured from the spread of the Doppler spectrum, which is extracted using joint spectral and time domain signal processing. At low OCT signal-to-noise ratio (SNR), the method provides more accurate vibration amplitude measurements than the currently used phasesensitive method. For measurements performed on a mirror at OCT SNR = 5 dB, our method introduces 20% using the phasesensitive method. We present elastograms of a tissue-mimicking phantom and excised porcine tissue that demonstrate improvements, including a 50% increase in the depth range of reliable vibration amplitude measurement.
We present a new approach to optical coherence elastography (OCE), which probes the local elastic properties of tissue by using optical coherence tomography to measure the effect of an applied stimulus in the audio frequency range. We describe the approach, based on analysis of the Bessel frequency spectrum of the interferometric signal detected from scatterers undergoing periodic motion in response to an applied stimulus. We present quantitative results of sub-micron excitation at 820 Hz in a layered phantom and the first such measurements in human skin in vivo.
We report the use of optical coherence tomography (OCT) to determine spatially localized optical attenuation coefficients of human axillary lymph nodes and their use to generate parametric images of lymphoid tissue. 3D-OCT images were obtained from excised lymph nodes and optical attenuation coefficients were extracted assuming a single scattering model of OCT. We present the measured attenuation coefficients for several tissue regions in benign and reactive lymph nodes, as identified by histopathology. We show parametric images of the measured attenuation coefficients as well as segmented images of tissue type based on thresholding of the attenuation coefficient values. Comparison to histology demonstrates the enhancement of contrast in parametric images relative to OCT images. This enhancement is a step towards the use of OCT for in situ assessment of lymph nodes.
Study Objectives: In patients with obstructive sleep apnea (OSA), the severity and frequency of respiratory events is increased in the supine body posture compared with the lateral recumbent posture. The mechanism responsible is not clear but may relate to the effect of posture on upper airway shape and size. This study compared the effect of body posture on upper airway shape and size in individuals with OSA with control subjects matched for age, BMI, and gender. Participants: 11 males with OSA and 11 age- and BMI-matched male control subjects. Results: Anatomical optical coherence tomography was used to scan the upper airway of all subjects while awake and breathing quietly, initially when supine, and then in the lateral recumbent posture. A standard head, neck, and tongue position was maintained during scanning. Airway cross-sectional area (CSA) and anteroposterior (A-P) and lateral diameters were obtained in the oropharyngeal and velopharyngeal regions in both postures. A-P to lateral diameter ratios provided an index of regional airway shape. In equivalent postures, the ratio of A-P to lateral diameter in the velopharynx was similar in OSA and control subjects. In both groups, this ratio was significantly less for the supine than for the lateral recumbent posture. CSA was smaller in OSA subjects than in controls but was unaffected by posture. Conclusions: The upper airway changes from a more transversely oriented elliptical shape when supine to a more circular shape when in the lateral recumbent posture but without altering CSA. Increased circularity decreases propensity to tube collapse and may account for the postural dependency of OSA.
We propose a passive optical network for customer access based on a hybrid coherence multiplexing/coarse wavelength-division multiplexing (WDM) technique. Coherence multiplexing provides asynchronous multichannel transmission, and coarse WDM provides bidirectional transmission. A costshared superfluorescent source is used for downstream transmission, and inexpensive light-emitting diodes are employed for upstream transmission. Asynchronous two channel transmission at 40 Mb/s per channel is demonstrated for both upstream and downstream. Our experiment indicates that upstream and downstream aggregate bit rates of 640 Mb/s are feasible based on current commercially available components.
We present an automated technique to detect and quantify damage to biological tissue by sensing changes in the tissue's optical birefringence. Birefringence is a property of many types of tissue, which decreases with damage. Using a polarisation-sensitive optical coherence tomography scanner, the method first acquires a 3D scan of the area of tissue under analysis. By calculating the birefringence at each location on the surface of the tissue, we build a 2D image indicative of the biological microstructure, with areas of abnormally low birefringence indicating tissue damage. The technique is demonstrated using a model of localised thermal damage on porcine tendon. The resulting birefringence images are validated against a histological gold standard, showing strong correspondence between areas of low and high birefringence, and areas of damaged and undamaged tissue respectively.
We present theoretical calculations, based on a random phasor sum model, which show that the optical coherence tomography speckle contrast ratio is dependent on the local density of scattering particles in a sample, provided that the effective number of scatterers in the probed volume is less than about five. We confirm these theoretical predictions experimentally, using suspensions of microspheres in water. The observed contrast ratios vary in value from the Rayleigh limit of 0.52 to in excess of 2, suggesting that the contrast ratio could be useful in optical coherence tomography, particularly when imaging in ultrahigh-resolution regimes.
Optical coherence elastography (OCE) is an emerging imaging technique that probes microscale mechanical contrast in tissues with the potential to differentiate healthy and malignant tissues. However, conventional OCE techniques are limited to imaging the first 1 to 2 mmof tissue in depth. We demonstrate, for the first time, OCE measurements deep within human tissues using needle OCE, extending the potential of OCE as a surgical guidance tool. We use needle OCE to detect tissue interfaces based on mechanical contrast in both normal and malignant breast tissues in freshly excised human mastectomy samples, as validated against histopathology. Further, we demonstrate the feasibility of in situ measurements 4 cm from the tissue surface using ultrasound guidance of the OCE needle probe. With further refinement, our method may potentially aid in accurate detection of the boundary of the tumor to help ensure full removal of all malignant tissues, which is critical to the success of breast-conserving surgery.
Objective: The anastomosis of peripheral nerves is a demanding procedure that has potential complications due to foreign body reactions elicited by sutures. In this study, the sutureless in vivo anastomosis of rat tibial nerves was successfully performed, using for the first time a chitosan-based laser-activated adhesive. The nerve thermal damage caused by the laser irradiation was quantitatively assessed. Materials and Methods: A novel adhesive composed of chitosan, indocyanine green, acetic acid, and water, was fabricated in thin sheets. Its adhesive strength was tested in vitro by bonding strips (surface area 2, thickness 2), using a fiber-coupled diode laser (n = 13). The tensile strength of the adhesive/tissue bonds was measured after tissue repair. The chitosan adhesive was then used to perform sutureless anastomosis of tibial nerves in vivo (n = 6). Adhesive strips were also bonded in vivo onto intact rat sciatic nerves (n = 6) in order to quantitatively assess, by counting myelinated axons, the thermal damage induced by the laser. Results: The adhesive bonded well to tissue with a tensile strength of 12.5 ± 2.6 KPa (mean ± SD; n = 13). The in vivo anastomosed nerves were in continuity 3 d after surgery. Axon counting showed the number and morphology of myelinated axons were normal proximally (
Transbronchial needle aspiration (TBNA) of small lesions or lymph nodes in the lung may result in nondiagnostic tissue samples. We demonstrate the integration of an optical coherence tomography (OCT) probe into a 19-gauge flexible needle for lung tissue aspiration. This probe allows simultaneous visualization and aspiration of the tissue. By eliminating the need for insertion and withdrawal of a separate imaging probe, this integrated design minimizes the risk of dislodging the needle from the lesion prior to aspiration and may facilitate more accurate placement of the needle. Results from in situ imaging in a sheep lung show clear distinction between solid tissue and two typical constituents of nondiagnostic samples (adipose and lung parenchyma). Clinical translation of this OCT-guided aspiration needle holds promise for improving the diagnostic yield of TBNA.
We evaluate the trade-off in optical bandwidth between signal-to-excess optical noise ratio and dispersion penalty in Gbit/s spectrum-sliced WDM systems. Furthermore, we demonstrate for the first time that, apart from producing intersymbol interference, dispersion also increases the excess optical noise.
Quantitative elasticity imaging seeks to retrieve spatial maps of elastic moduli of tissue. Unlike strain, which is commonly imaged in compression elastography, elastic moduli are intrinsic properties of tissue, and therefore, this approach reconstructs images that are largely operator and system independent, enabling objective, longitudinal, and multisite diagnoses. Recently, novel quantitative elasticity imaging approaches to compression elastography have been developed. These methods use a calibration layer with known mechanical properties to sense the stress at the tissue surface, which combined with strain, is used to estimate the tissue's elastic moduli by assuming homogeneity in the stress field. However, this assumption is violated in mechanically heterogeneous samples. We present a more general approach to quantitative elasticity imaging that overcomes this limitation through an efficient iterative solution of the inverse elasticity problem using adjoint elasticity equations. We present solutions for linear elastic, isotropic, and incompressible solids; however, this method can be employed for more complex mechanical models. We retrieve the spatial distribution of shear modulus for a tissue-simulating phantom and a tissue sample. This is the first time, to our knowledge, that the iterative solution of the inverse elasticity problem has been implemented on experimentally acquired compression optical coherence elastography data.
Three-dimensional optical coherence tomography (3D-OCT) was used to image the structure and pathology of skeletal muscle tissue from the treadmill-exercised mdx mouse model of human Duchenne muscular dystrophy. Optical coherence tomography (OCT) images of excised muscle samples were compared with coregistered hematoxylin and eosin-stained and Evans blue dye fluorescence histology. We show, for the first time, structural 3D-OCT images of skeletal muscle dystropathology well correlated with co-located histology. OCT could identify morphological features of interest and necrotic lesions within the muscle tissue samples based on intrinsic optical contrast. These findings demonstrate the utility of 3D-OCT for the evaluation of small-animal skeletal muscle morphology and pathology, particularly for studies of mouse models of muscular dystrophy.
We present a graphics processing unit (GPU)-accelerated optical coherence elastography (OCE) system capable of generating strain images (elastograms) of soft tissue at near video-rates. The system implements phase-sensitive compression OCE using a pipeline of GPU kernel functions to enable a highly parallel implementation of OCE processing using the OpenCL framework. Developed on a commercial-grade GPU and desktop computer, the system achieves a processing rate of 21 elastograms per second at an image size of 960 X 400 pixels, enabling high-rate visualization during acquisition. The system is demonstrated on both tissue-simulating phantoms and fresh ex vivo mouse muscle. To the best of our knowledge, this is the first implementation of near video-rate OCE and the fastest reported OCE processing rate, enabling, for the first time, a system capable of computing and displaying OCE elastograms interactively during acquisition. This advance provides new opportunities for medical imaging of soft tissue stiffness using optical methods.
A > 100mW source of broadband ASE with exceptionally flat spectral density and 22nm bandwidth is presented. The source is ideally suited for spectrum-sliced WDM systems. Data transmission at 622Mbit/s using a spectraily-sliced channel tuned over a 15nm range is demonstrated.
After more than a decade of research, optical coherence tomography is in the early phases of establishing a niche as a medical imaging technology for routine clinical use. Far from tailing off, however, research activity is, if anything, on the increase. In this paper, we briefly and selectively review the current state-of-the-art in the more prominent areas of activity in the technology and its application. One is inevitably drawn to the conclusion that optical coherence tomography has much more to offer clinical practice than has yet been transferred.
Gong Peijun, Yu Dao-Yi, Wang Qiang, Yu Paula K., Karnowski Karol, Heisler Morgan, Francke Ashley, An Dong, Sarunic Marinko V., Sampson David D. (2018) Label-free volumetric imaging of conjunctival collecting lymphatics ex vivo by optical coherence tomography lymphangiography, Journal of Biophotonics 11 (8) e201800070
We employ optical coherence tomography (OCT) and optical coherence microscopy (OCM) to study conjunctival lymphatics in porcine eyes ex vivo. This study is a precursor to the development of in vivo imaging of the collecting lymphatics for potentially guiding and monitoring glaucoma filtration surgery. OCT scans at 1300 nm and higher?resolution OCM scans at 785 nm reveal the lymphatic vessels via their optical transparency. Equivalent signal characteristics are also observed from blood vessels largely free of blood (and devoid of flow) in the ex vivo conjunctiva. In our lymphangiography, vessel networks were segmented by compensating the depth attenuation in the volumetric OCT/OCM signal, projecting the minimum intensity in two dimensions and thresholding to generate a three?dimensional vessel volume. Vessel segmentation from multiple locations of a range of porcine eyes (n = 21) enables visualization of the vessel networks and indicates the varying spatial distribution of patent lymphatics. Such visualization provides a new tool to investigate conjunctival vessels in tissue ex vivo without need for histological tissue processing and a valuable reference on vessel morphology for the in vivo label?free imaging studies of lymphatics to follow.
It is challenging to recover local optic axis orientation from samples probed with
fiber-based polarization-sensitive optical coherence tomography (PS-OCT). In addition to the
effect of preceding tissue layers, the transmission through fiber and system elements, and
imperfect system alignment, need to be compensated. Here, we present a method to retrieve
the required correction factors from measurements with depth-multiplexed PS-OCT, which
accurately measures the full Jones matrix. The correction considers both retardation and
diattenuation and is applied in the wavenumber domain, preserving the axial resolution of the
system. The robustness of the method is validated by measuring a birefringence phantom with
a misaligned system. Imaging ex-vivo lamb trachea and human bronchus demonstrates the
utility of reconstructing the local optic axis orientation to assess smooth muscle, which is
expected to be useful in the assessment of airway smooth muscle thickness in asthma,
amongst other fiber-based applications.
Li Jiawen, Fejes Peter, Lorenser Dirk, Quirk Bryden C., Noble Peter B., Kirk Rodney W., Orth Antony, Wood Fiona M., Gibson Brant C., Sampson David D., McLaughlin Robert A. (2018) Two-photon polymerisation 3D printed freeform micro-optics for optical coherence tomography fibre probes, Scientific Reports 8 (1) 14789 pp. 1-9
Miniaturised optical coherence tomography (OCT) fibre-optic probes have enabled high-resolution cross-sectional imaging deep within the body. However, existing OCT fibre-optic probe fabrication methods cannot generate miniaturised freeform optics, which limits our ability to fabricate probes with both complex optical function and dimensions comparable to the optical fibre diameter. Recently, major advances in two-photon direct laser writing have enabled 3D printing of arbitrary three-dimensional micro/nanostructures with a surface roughness acceptable for optical applications. Here, we demonstrate the feasibility of 3D printing of OCT probes. We evaluate the capability of this method based on a series of characterisation experiments. We report fabrication of a micro-optic containing an off-axis paraboloidal total internal reflecting surface, its integration as part of a common-path OCT probe, and demonstrate proof-of-principle imaging of biological samples.
It is widely accepted that accurate mechanical properties of three-dimensional soft tissues and cellular samples are not available on the microscale. Current methods based on optical coherence elastography can measure displacements at the necessary resolution, and over the volumes required for this task. However, in converting this data to maps of elastic properties, they often impose assumptions regarding homogeneity in stress or elastic properties that are violated in most realistic scenarios. Here, we introduce novel, rigorous, and computationally efficient inverse problem techniques that do not make these assumptions, to realize quantitative volumetric elasticity imaging on the microscale. Specifically, we iteratively solve the three-dimensional elasticity inverse problem using displacement maps obtained from compression optical coherence elastography. This is made computationally feasible with adaptive mesh refinement and domain decomposition methods. By employing a transparent, compliant surface layer with known shear modulus as a reference for the measurement, absolute shear modulus values are produced within a millimeter-scale sample volume. We demonstrate the method on phantoms, on a breast cancer sample ex vivo, and on human skin in vivo. Quantitative elastography on this length scale will find wide application in cell biology, tissue engineering and medicine.
We present a new optical coherence tomography (OCT) angiography method for imaging tissue microvasculature in vivo based on the characteristic frequency-domain flow signature in a short time series of a single voxel. The angiography signal is generated by Fourier transforming the OCT signal time series from a given voxel in multiple acquisitions and computing the average magnitude of non-zero (high-pass) frequency components. Larger temporal variations of the OCT signal caused by blood flow result in higher values of the average magnitude in the frequency domain compared to those from static tissue. Weighting of the signal by the inverse of the zero-frequency component (i.e., the sum of the OCT signal time series) improves vessel contrast in flow regions of low OCT signal. The method is demonstrated on a fabricated flow phantom and on human skin in vivo and, at only 5 time points per voxel, shows enhanced vessel contrast in comparison to conventional correlation mapping/speckle decorrelation and speckle variance methods.
Stromal collagen organization has been identified as a potential prognostic indicator in a variety of cancers and other diseases accompanied by fibrosis. Changes in the connective tissue are increasingly considered for grading dysplasia and progress of oral squamous cell carcinoma, investigated mainly ex vivo by histopathology. In this study, polarization-sensitive optical coherence tomography (PS-OCT) with local phase retardation imaging is used for the first time to visualize depth-resolved (i.e., local) birefringence of healthy human oral mucosa in vivo. Depth-resolved birefringence is shown to reveal the expected local collagen organization. To demonstrate proof-of-principle, 3D image stacks were acquired at labial and lingual locations of the oral mucosa, chosen as those most commonly affected by cancerous alterations. To enable an intuitive evaluation of the birefringence images suitable for clinical application, color depth-encoded en-face projections were generated. Compared to en-face views of intensity or conventional cumulative phase retardation, we show that this novel approach offers improved visualization of the mucosal connective tissue layer in general, and reveals the collagen fiber architecture in particular. This study provides the basis for future prospective pathological and comparative in vivo studies non-invasively assessing stromal changes in conspicuous and cancerous oral lesions at different stages.
Yeow Yen Ling, Kotamraju Venkata Ramana, Wang Xiao, Chopra Meenu, Azme Nasibah, Wu Jiansha, Schoep Tobias D, Delaney Derek S, Feindel Kirk, Li Ji, Kennedy Kelsey M, Allen Wes M, Kennedy Brendan F, Larma Irma, Sampson David D, Mahakian Lisa M, Fite Brett Z, Zhang Hua, Friman Tomas, Mann Aman P, Aziz Farah A, Kumarasinghe M Priyanthi, Johansson Mikael, Ee Hooi C, Yeoh George, Mou Lingjun, Ferrara Katherine W, Billiran Hector, Ganss Ruth, Ruoslahti Erkki, Hamzah Juliana (2019) Immune?mediated ECM depletion improves tumour perfusion and payload delivery, EMBO Molecular Medicine e10923 pp. 1-20
Wiley Open Access
High extracellular matrix (ECM) content in solid cancers impairs tumour perfusion and thus access of imaging and therapeutic agents. We have devised a new approach to degrade tumour ECM, which improves uptake of circulating compounds. We target the immune?modulating cytokine, tumour necrosis factor alpha (TNF±), to tumours using a newly discovered peptide ligand referred to as CSG. This peptide binds to laminin?nidogen complexes in the ECM of mouse and human carcinomas with little or no peptide detected in normal tissues, and it selectively delivers a recombinant TNF±?CSG fusion protein to tumour ECM in tumour?bearing mice. Intravenously injected TNF±?CSG triggered robust immune cell infiltration in mouse tumours, particularly in the ECM?rich zones. The immune cell influx was accompanied by extensive ECM degradation, reduction in tumour stiffness, dilation of tumour blood vessels, improved perfusion and greater intratumoral uptake of the contrast agents gadoteridol and iron oxide nanoparticles. Suppressed tumour growth and prolonged survival of tumour?bearing mice were observed. These effects were attainable without the usually severe toxic side effects of TNF±.
The organization of fibrillar tissue on the micrometer scale carries direct implications
for health and disease but remains difficult to assess in vivo. Polarization-sensitive optical
coherence tomography measures birefringence, which relates to the microscopic arrangement
of fibrillar tissue components. Here, we demonstrate a critical improvement in leveraging this
contrast mechanism by employing the improved spatial resolution of focus-extended optical
coherence microscopy (1.4 ¼m axially in air and 1.6 ¼m laterally, over more than 70 ¼m depth of
field). Vectorial birefringence imaging of sheep cornea ex vivo reveals its lamellar organization
into thin sections
Significance: Optical coherence tomography (OCT) provides cross-sectional and volumetric
images of backscattering from biological tissue that reveal the tissue morphology. The strength
of the scattering, characterized by an attenuation coefficient, represents an alternative and complementary
tissue optical property, which can be characterized by parametric imaging of the
OCT attenuation coefficient. Over the last 15 years, a multitude of studies have been reported
seeking to advance methods to determine the OCT attenuation coefficient and developing them
toward clinical applications.
Aim: Our review provides an overview of the main models and methods, their assumptions
and applicability, together with a survey of preclinical and clinical demonstrations and their
Results: The use of the attenuation coefficient, particularly when presented in the form of parametric
en face images, is shown to be applicable in various medical fields. Most studies show the
promise of the OCT attenuation coefficient in differentiating between tissues of clinical interest
but vary widely in approach.
Conclusions: As a future step, a consensus on the model and method used for the determination
of the attenuation coefficient is an important precursor to large-scale studies. With our review,
we hope to provide a basis for discussion toward establishing this consensus.
We show that polarization-sensitive optical coherence tomography angiography (PS-OCTA) based on full Jones matrix assessment of speckle decorrelation offers improved contrast and depth of vessel imaging over conventional OCTA. We determine how best to combine the individual Jones matrix elements and compare the resulting image quality to that of a conventional OCT scanner by co-locating and imaging the same skin locations with closely matched scanning setups. Vessel projection images from finger and forearm skin demonstrate the benefits of Jones matrix-based PS-OCTA. Our study provides a promising starting point and a useful reference for future pre-clinical and clinical applications of Jones matrix-based PS-OCTA.