A review of some recent developments in polarization-sensitive optical imaging techniques for the study of articular cartilage

This article reviews recent developments in the optical imaging of articular cartilage using polarized-light methods, with an emphasis on tools that could be of use in tissue engineering approaches to treatment. Both second-harmonic generation microscopy and polarization-sensitive optical coherence tomography are described and their potential role in the treatment of cartilage disorders such as osteoarthritis is suggested. Key results are reviewed and future developments are discussed

Novel optical imaging technique to determine the 3-D orientation of collagen fibers in cartilage: variable-incidence angle polarization-sensitive optical coherence tomography

Objective: To investigate a novel optical method to determine the three dimensional (3-D) structure of articular cartilage collagen non-destructively. Methods: Polarization-sensitive optical coherence tomography was used to determine the apparent optical birefringence of articular cartilage for a number of different illumination directions. A quantitative method based on the theory of light propagation in uniaxial crystalline materials was validated on equine flexor tendon. Qualitative maps of fiber polar and azimuthal orientation at sites on the posterior and anterior segments of the equine third metacarpophalangeal (fetlock) joint were produced, and the azimuthal orientations compared with data from a split-line experiment. Results: Polar and azimuthal angles of cut flexor tendon broadly agreed with the nominal values but suggested that the accuracy was limited by our method of determining the apparent birefringence. On intact equine fetlock joints we found a non-zero polar tilt that changed in direction at various points along the apex, moving from the sagittal ridge outwards. The azimuthal orientation changes from being parallel to the sagittal ridge in the posterior region to being inclined to the ridge in the anterior region. This broadly agrees with split-line data for the anterior region but differs in the posterior region, possibly reflecting depth-dependent orientation changes. Conclusion: General quantitative agreement was found between our method and histology in validation experiments. Qualitative results for cartilage suggest a complicated 3-D structure that warrants further study. There is potential to develop this approach into a tool that can provide depth-resolved information on collagen orientation in near real-time, non-destructively and in vivo. (c) 2008 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved

Ghosting artifact reduction of polarization sensitive optical coherence tomography images through wavelet-FFT filtering

Undesirable cross-coupling between polarisation-maintaining (PM) fibers can result in detrimental ghost artefacts within polarisation sensitive optical coherence tomography (PS-OCT) images. Such artefacts combine with coherence noise stripes (originating from Fresnel reflections of optical components), complex-conjugate derived mirror-images and further irregular autocorrelation terms originating from the sample. Together, these artefacts can severely degrade the detected images, making quantitative measurements of the tissue birefringence challenging to perform. In this work, we utilize the recently presented wavelet-FFT filter1 to efficiently suppress these imaging artefacts entirely through post-processing. While the original algorithm was designed to suppress one-dimensional stripe artefacts, we extend this methodology to also facilitate removal of artefacts following a duplicate or inverse (mirror) profile to that of the skin surface. This process does not require any hardware modification of the system and can be applied retroactively to previously acquired OCT images. The performance of this methodology is evaluated by processing artefact-corrupted PS-OCT images of skin consisting of simultaneously detected horizontal and vertical polarized light. The resulting images are used to calculate a phase retardance map within the skin, the profile of which is indicative of localized birefringence. Artefacts in the resulting processed PSOCT images were notably attenuated compared to the unprocessed raw-data, with minimal degradation to the underlying phase retardation information. This should improve the reliability of curve-fitting for measurements of depth-resolved birefringence

Morphological parametric mapping of 21 skin sites throughout the body using optical coherence tomography

Background Changes in body posture cause changes in morphological properties at different skin sites. Although previous studies have reported the thickness of the skin, the details of the postures are not generally given. This paper presents the effect of a change in posture on parameters such as thickness and surface roughness in 21 load-bearing and non-load-bearing sites. Materials and methods A total of 12 volunteers (8 males and 4 females) were selected in an age group of 18–35 years and of Fitzpatrick skin type I-III. Images were captured using a clinically-approved VivoSight® optical coherence tomography system and analysed using an algorithm provided by Michelson Diagnostics. Results Overextension (extending joints to full capacity) resulted in changes to thickness, roughness and undulation of the skin around the body. Discussion and conclusion The load-bearing regions have thicker skin compared to non-load-bearing sites. This is the first time that undulation topography of the stratum corneum–stratum lucidum and the dermal–epidermal junction layers have been measured and reported using statistical values such as Ra. The data presented could help to define new skin layer models and to determine the variability of the skin around the body and between participants

A new mode of contrast in biological second harmonic generation microscopy

Enhanced image contrast in biological second harmonic imaging microscopy (SHIM) has previously been reported via quantitative assessments of forward- to epi-generated signal intensity ratio and by polarization analysis. Here we demonstrate a new form of contrast: the material-specific, wavelength-dependence of epi-generated second harmonic generation (SHG) excitation efficiency, and discriminate collagen and myosin by ratiometric epi-generated SHG images at 920 nm and 860 nm. Collagen shows increased SHG intensity at 920 nm, while little difference is detected between the two for myosin; allowing SHIM to characterize different SHG-generating components within a complex biological sample. We propose that momentum-space mapping of the second-order non-linear structure factor is the source of this contrast and develop a model for the forward and epi-generated SHG wavelength-dependence. Our model demonstrates that even very small changes in the assumed material fibrillar structure can produce large changes in the wavelength-dependency of epi-generated SHG. However, in the case of forward SHG, although the same changes impact upon absolute intensity at a given wavelength, they have very little effect on wavelength-dependency beyond the expected monotonic fall. We also propose that this difference between forward and epi-generated SHG provides an explanation for many of the wavelength-dependency discrepancies in the published literature

Superparamagnetic graphene quantum dot as a dual-modality contrast agent for confocal fluorescence microscopy and magnetomotive optical coherence tomography

A magnetic graphene quantum dot (MGQD) nanoparticle, synthesized by hydrothermally reducing and cutting graphene oxide-iron oxide sheet, was demonstrated to possess the capabilities of simultaneous confocal fluorescence and magnetomotive OCT imaging. This MGQD shows low toxicity, significant tunable blue fluorescence and superparamagnetism, which can thus be used as a dual-modality contrast agent for confocal fluorescence microscopy (CFM) and magnetomotive OCT (MMOCT). The feasibility of applying MGQD as a tracer of cells is shown by imaging and visualizing MGQD labelled cells using CFM and our in-house MMOCT. Since MMOCT and CFM can offer anatomical structure and intracellular details respectively, the MGQD for cell tracking could provide a more comprehensive diagnosis. This article is protected by copyright. All rights reserved

Visible-light optical coherence tomography platform for the characterization of the skin barrier

We demonstrate a free-space, trolley-mountable Fourier domain visible-light optical coherence tomography (OCT) system for studying the stratum corneum in non-palmar human skin. An axial resolution of 1 µm in tissue and at least −75 dB sensitivity have been achieved. High-quality B-scans, containing 1600 A-scans, are acquired at a rate of 39 Hz. Images from the dorsal hand, ventral wrist and ventral forearm areas are obtained, with a clearly resolved stratum corneum layer (typically 5–15 µm thick) presenting as a hypoechogenic dark layer below the bright entrance signal, similar to that found in palmar skin with traditional OCT systems. We find that the appearance of the stratum corneum layer strongly depends on its water content, becoming brighter after occlusive hydration

Sub-clinical assessment of atopic dermatitis severity using angiographic optical coherence tomography

Measurement of sub-clinical atopic dermatitis (AD) is important for determining how long therapies should be continued after clinical clearance of visible AD lesions. An important biomarker of sub-clinical AD is epidermal hypertrophy, the structural measures of which often make optical coherence tomography (OCT) challenging due to the lack of a clearly delineated dermal-epidermal junction in AD patients. Alternatively, angiographic OCT measurements of vascular depth and morphology may represent a robust biomarker for quantifying the severity of clinical and sub-clinical AD. To investigate this, angiographic data sets were acquired from 32 patients with a range of AD severities. Deeper vascular layers within skin were found to correlate with increasing clinical severity. Furthermore, for AD patients exhibiting no clinical symptoms, the superficial plexus depth was found to be significantly deeper than healthy patients at both the elbow (p = 0.04) and knee (p < 0.001), suggesting that sub-clinical changes in severity can be detected. Furthermore, the morphology of vessels appeared altered in patients with severe AD, with significantly different vessel diameter, length, density and fractal dimension. These metrics provide valuable insight into the sub-clinical severity of the condition, allowing the effects of treatments to be monitored past the point of clinical remission

Sensitivity Advantage of QCL Tunable-Laser Mid-Infrared Spectroscopy over FTIR Spectroscopy

Interest in mid-infrared spectroscopy instrumentation beyond classical FTIR using a thermal light source has increased dramatically in recent years. Synchrotron, supercontinuum, and external-cavity quantum cascade laser light sources are emerging as viable alternatives to the traditional thermal black-body emitter (Globar), especially for remote interrogation of samples ("stand-off" detection) and for hyperspectral imaging at diffraction-limited spatial resolution ("microspectroscopy"). It is thus timely to rigorously consider the relative merits of these different light sources for such applications. We study the theoretical maximum achievable signal-to-noise ratio (SNR) of FTIR using synchrotron or supercontinuum light vs. that of a tunable quantum cascade laser, by reinterpreting an important result that is well known in near-infrared optical coherence tomography imaging. We rigorously show that mid-infrared spectra can be acquired up to 1000 times faster - using the same detected light intensity, the same detector noise level, and without loss of SNR - using the tunable quantum cascade laser as compared with the FTIR approach using synchrotron or supercontinuum light. We experimentally demonstrate the effect using a novel, rapidly tunable quantum cascade laser that acquires spectra at rates of up to 400 per second. We also estimate the maximum potential spectral acquisition rate of our prototype system to be 100,000 per second

New Non-invasive Techniques to Quantify Skin Surface Strain and Sub-surface Layer Deformation of Finger-pad during Sliding

Studies on the variation of skin properties with gender, age and anatomical region, with regards to interaction with different materials have resulted in significant research output. Investigations on skin surface strain and sub-surface layer deformation during sliding, however, have not received as much attention. This novel study uses two non-invasive techniques, optical coherence tomography (OCT) and digital image correlation (DIC), to measure properties of the index finger of a 25. year old female when under normal and shear loading. Measurements were taken during static, and for the first time, dynamic phases.It was observed that the number of ridges in contact with a Quartz glass surface, observed under OCT, reduced when the finger started sliding. The sliding also resulted in deformation at the stratum corneum junction. The surface strain, analysed using DIC was higher nearer to the distal interphalangeal joint compared to the fingertip. This newly developed approach provides a powerful and non-invasive method to study the structural changes of finger-pad skin during loading and sliding. This approach can now be repeated at different anatomical locations for medical, bioengineering and consumer applications