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 -75â dB sensitivity have been achieved. High-quality B-scans, containing 1600 A-scans, are acquired at a rate of 39â 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.
Crisaborole 2% ointment is a non-steroidal treatment for mild-moderate atopic dermatitis (AD) and may produce fewer adverse effects than topical corticosteroids (TCS). We used PS-OCT to quantify dermal collagen at baseline and after 29 days of treatment with crisaborole and betamethasone valerate (BMV), in 32 subjects. PS-OCT detected a mean increase 1 × 10-6, 95% CI (6.3, 1.37) × 10-6 in dermal birefringence following TCS use (p < 0.0001, ad-hoc, not powered), whereas a change of -4 × 10-6, 95% CI (-32, 24) × 10-6 was detected for crisaborole (p = 0.77, ad-hoc, not powered). These results could suggest a differential effect on dermal collagen between the two compounds. PS-OCT may thus find an important role in safety assessment of novel AD treatment' and larger trials are warranted.
We report a continuous wave room temperature quantum cascade laser operating in an external cavity in the Littrow configuration with a 10-facet polygon mirror rotating at 24,000 RPM. The quantum cascade laser emission is swept across â¼1520 - 1625 cm-1 wavenumber range in less than â¼45 µs with a sweep repetition rate of 4 kHz. The measured maximum output power at the laser gain maximum, 15°C and 0.86 A driving current is â¼90 mW; the estimated average output power across the 45 µs wavenumber sweep is â¼50 mW. Through its sweep, the laser emits on the sequential Fabry-Perot longitudinal modes of the laser chip cavity with the mode separation of â¼0.5 cm-1. The linewidth of the emitting modes is less than â¼0.05 cm-1. Spectral measurements of the infrared absorption features of a 10 µm thick layer of acetophenone and water vapor in the air have demonstrated the capability of obtaining spectral data in less than 45 µs.
Stretch marks or striae distensae (SD) cause emotional distress and negatively affect the psychological well-being of patients. We investigate and compare two methods for quantifying the severity of SD: visual scoring of images captured using a clinical visible-light dermatological camera (C-Cube, Pixience Inc) and measuring the local birefringence of skin using polarization-sensitive optical coherence tomography (PS-OCT). Data on skin visually affected by SD and visually normal skin were collected from 19 human volunteers. Our results show a weak correlation between visual scores of the C-Cube images and the birefringence values obtained from the PS-OCT system. SD datasets have a significantly larger birefringence values compared to visually normal datasets.
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.
Skin , Tomography, Optical Coherence , Adolescent , Adult , Epidermis , Female , Humans , Male , Young Adult
By measuring the phase retardance of a cervical extracellular matrix, our in-house polarization-sensitive optical coherence tomography (PS-OCT) was shown to be capable of (1) mapping the distribution of collagen fibers in the non-gravid cervix, (2) accurately determining birefringence, and (3) measuring the distinctive depolarization of the cervical tissue. A conical beam scan strategy was also employed to explore the 3D orientation of the collagen fibers in the cervix by interrogating the samples with an incident light at 45° and successive azimuthal rotations of 0-360°. Our results confirmed previous observations by X-ray diffraction, suggesting that in the non-gravid human cervix collagen fibers adjacent to the endocervical canal and in the outermost areas tend to arrange in a longitudinal fashion whereas in the middle area they are oriented circumferentially. PS-OCT can assess the microstructure of the human cervical collagen in vitro and holds the potential to help us better understand cervical remodeling prior to birth pending the development of an in vivo probe.
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 optical coherence tomography (MMOCT) 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 MMOCT. The feasibility of applying MGQD as a tracer of cells is shown by imaging and visualizing MGQD labeled 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.
Contrast Media , Graphite/chemistry , Magnets/chemistry , Microscopy, Confocal/methods , Microscopy, Fluorescence/methods , Quantum Dots , Tomography, Optical Coherence/methods , 3T3 Cells , Animals , Mice
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.
Subcutaneously implanted experimental tumors in mice are commonly used in cancer research. Despite their superficial location, they remain a challenge to image non-invasively at sufficient spatial resolution for microvascular studies. Here we evaluate the capabilities of optical coherence tomography (OCT) angiography for imaging such tumors directly through the murine skin in-vivo. Data sets were collected from mouse tumors derived from fibrosarcoma cells genetically engineered to express only single splice variant isoforms of vascular endothelial growth factor A (VEGF); either VEGF120 or VEGF188 (fs120 and fs188 tumors respectively). Measured vessel diameter was found to be significantly (p<0.001) higher for fs120 tumors (60.7 ± 4.9µm) compared to fs188 tumors (45.0 ± 4.0µm). The fs120 tumors also displayed significantly higher vessel tortuosity, fractal dimension and density. The ability to differentiate between tumor types with OCT suggests that the visible abnormal vasculature is representative of the tumor microcirculation, providing a robust, non-invasive method for observing the longitudinal dynamics of the subcutaneous tumor microcirculation.
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.
Second Harmonic Generation Microscopy/methods , Collagen/metabolism , Dermis/cytology , Fibroblasts/cytology , Fibroblasts/metabolism , Humans , Image Processing, Computer-Assisted
Stratum corneum and epidermal layers change in terms of thickness and roughness with gender, age and anatomical site. Knowledge of the mechanical and tribological properties of skin associated with these structural changes are needed to aid in the design of exoskeletons, prostheses, orthotics, body mounted sensors used for kinematics measurements and in optimum use of wearable on-body devices. In this case study, optical coherence tomography (OCT) and digital image correlation (DIC) were combined to determine skin surface strain and sub-surface deformation behaviour of the volar forearm due to natural tissue stretching. The thickness of the epidermis together with geometry changes of the dermal-epidermal junction boundary were calculated during change in the arm angle, from flexion (90°) to full extension (180°). This posture change caused an increase in skin surface Lagrange strain, typically by 25% which induced considerable morphological changes in the upper skin layers evidenced by reduction of epidermal layer thickness (20%), flattening of the dermal-epidermal junction undulation (45-50% reduction of flatness being expressed as Ra and Rz roughness profile height change) and reduction of skin surface roughness Ra and Rz (40-50%). The newly developed method, DIC combined with OCT imaging, is a powerful, fast and non-invasive methodology to study structural skin changes in real time and the tissue response provoked by mechanical loading or stretching.
Dermis/physiology , Epidermis/physiology , Stress, Mechanical , Forearm , Humans , Skin , Tomography, Optical Coherence
Connective tissues such as articular cartilage have been the subject of study using novel optical techniques almost since the invention of polarized light microscopy (PLM). Early studies of polarized light micrographs were the main evidential basis for the establishment of quantitative models of articular cartilage collagen structure by Benninghoff and others. Even now, state of the art optical techniques including quantitative polarized light microscopy (qPLM), optical coherence tomography (OCT), polarization-sensitive optical coherence tomography (PS-OCT), second harmonic generation (SHG) microscopy, Fourier-transform infrared (FTIR) microscopy, Raman and optical hyperspectral reflectance and fluorescence imaging are providing new insights into articular cartilage structure from the nanoscale through to the mesoscale. New insights are promised by emerging modalities such as optical elastography. This short review highlights some key recent results from modern optical techniques.
We report on a new articular cartilage imaging technique with potential for clinical arthroscopic use, by supplementing the variable-incidence-angle polarization-sensitive optical coherence tomography method previously developed by us with a conical beam scan protocol. The technique is validated on bovine tendon by comparing experimental data with simulated data generated using the extended Jones matrix calculus. A unique capability of this new optical technique is that it can locate the "brushing direction" of collagen fibers in articular cartilage, which is structural information that extends beyond established methods such as split-line photography or birefringent fast-axis measurement in that it is uniquely defined over the full azimuthal-angle range of (-π, + π). The mapping of this direction over the cartilage surface may offer insights into the optimal design of tissue-engineering scaffolds for cartilage repair.
The biological and mechanical function of connective tissues is largely determined by controlled cellular alignment and therefore it seems appropriate that tissue-engineered constructs should be architecturally similar to the in vivo tissue targeted for repair or replacement. Collagen organisation dictates the tensile properties of most tissues and so monitoring the deposition of cell-secreted collagen as the construct develops is essential for understanding tissue formation. In this study, electrospun fibres with a random or high degree of orientation, mimicking two types of tissue architecture found in the body, were used to culture human fibroblasts for controlling cell alignment. The minimally-invasive technique of second harmonic generation was used with the aim of monitoring and profiling the deposition and organisation of collagen at different construct depths over time while construct mechanical properties were also determined over the culture period. It was seen that scaffold fibre organisation affected cell migration and orientation up to 21 days which in turn had an effect on collagen organisation. Collagen in random fibrous constructs was deposited in alternating configurations at different depths however a high degree of organisation was observed throughout aligned fibrous constructs orientated in the scaffold fibre direction. Three-dimensional second harmonic generation images showed that deposited collagen was more uniformly distributed in random constructs but aligned constructs were more organised and had higher intensities. The tensile properties of all constructs increased with increasing collagen deposition and were ultimately dictated by collagen organisation. This study highlights the importance of scaffold architecture for controlling the development of well-organised tissue engineered constructs and the usefulness of second harmonic generation imaging for monitoring collagen maturation in a minimally invasive manner.
Collagen/chemistry , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Cells, Cultured , Humans
Optical coherence tomography (OCT) is a modern high resolution subsurface medical imaging technique. Herein we describe: (i) the synthesis of a thiophene-functionalized oligo(ethylene glycol) methacrylate (OEGMA)-based statistical copolymer, denoted poly(2TMOI-OEGMA); (ii) the preparation of sterically-stabilized polypyrrole (PPy) nanoparticles of approximately 60 nm diameter; (iii) the evaluation of these nanoparticles as a NIR-absorbing optical contrast agent for high-resolution OCT imaging. We show that poly(2TMOI-OEGMA)-stabilized PPy nanoparticles exhibit similar optical properties to poly(vinyl alcohol) (PVA)-stabilized PPy nanoparticles of comparable size prepared using commercially available PVA. Spectroscopic measurements and Mie calculations indicate that both types of PPy nanoparticles strongly absorb NIR radiation above 1000 nm, suggesting their potential use as OCT contrast agents. In vitro OCT studies indicate that both types of PPy nanoparticles reduce NIR backscattering within homogeneous intralipid tissue phantoms, offering almost identical contrast performance in this medium. However, PVA-stabilized PPy nanoparticles became colloidally unstable when dispersed in physiological buffer and immersed in a solid biotissue phantom and hence failed to generate a strong contrast effect. In contrast, the poly(2TMOI-OEGMA)-stabilized PPy nanoparticles remained well-dispersed and hence exhibited a strong rapid onset contrast effect within the biotissue phantom under identical physiological conditions. Ex vivo studies performed on excised chicken and porcine skin tissue demonstrated that topical administration of a low concentration of poly(2TMOI-OEGMA)-stabilized PPy nanoparticles rapidly enhances OCT image contrast in both cases, allowing key tissue features to be readily identified.
Biofouling/prevention & control , Diagnostic Imaging/methods , Nanoparticles/chemistry , Polymers/chemistry , Animals , Chickens , Contrast Media/chemistry , Emulsions/chemistry , Methacrylates/chemical synthesis , Microscopy, Electron, Transmission , Particle Size , Phantoms, Imaging , Phospholipids/chemistry , Photoelectron Spectroscopy , Polyethylene Glycols/chemical synthesis , Polymethacrylic Acids , Pyrroles/chemistry , Skin/drug effects , Skin/metabolism , Soybean Oil/chemistry , Spectrophotometry, Infrared , Swine , Tomography, Optical Coherence
We report on a new calibration technique that permits the accurate extraction of sample Jones matrix and hence fast-axis orientation by using fiber-based polarization-sensitive optical coherence tomography (PS-OCT) that is completely based on non-polarization-maintaining fiber such as SMF-28. In this technique, two quarter-wave plates (QWPs) are used to completely specify the parameters of the system fibers in the sample arm so that the Jones matrix of the sample can be determined directly. The device was validated on measurements of a QWP and an equine tendon sample by a single-mode fiber-based swept-source PS-OCT system.
Optical Phenomena , Tomography, Optical Coherence/methods , Animals , Calibration , Horses , Tendons/cytology
We demonstrate theoretically and experimentally that the phase retardance and relative optic-axis orientation of a sample can be calculated without prior knowledge of the actual value of the phase modulation amplitude when using a polarization-sensitive optical coherence tomography system based on continuous polarization modulation (CPM-PS-OCT). We also demonstrate that the sample Jones matrix can be calculated at any values of the phase modulation amplitude in a reasonable range depending on the system effective signal-to-noise ratio. This has fundamental importance for the development of clinical systems by simplifying the polarization modulator drive instrumentation and eliminating its calibration procedure. This was validated on measurements of a three-quarter waveplate and an equine tendon sample by a fiber-based swept-source CPM-PS-OCT system.
Tomography, Optical Coherence/methods , Animals , Calibration , Horses , Models, Theoretical , Reproducibility of Results , Signal-To-Noise Ratio , Tendons/chemistry
We report results to verify a theoretical framework to analyze the 3D depth-wise structural organization of collagen fibers in articular cartilage using polarization-sensitive optical coherence tomography. Apparent birefringence data obtained from multi-angle measurements using a time domain polarization-sensitive optical coherence tomography system has been compared with simulated data based on the extended Jones matrix calculus. Experimental data has been shown to agree with the lamellar model previously proposed for the cartilage microstructure based on scanning electron microscopy data. This tool could have potential application in mapping the collagen structural orientation information of cartilage non-invasively during arthroscopy.
Optical coherence tomography (OCT) is a noninvasive imaging methodology that is able to image tissue to depths of over 1 mm. Many epithelial conditions, such as melanoma and oral cancers, require an invasive biopsy for diagnosis. A noninvasive, real-time, point of care method of imaging depth-resolved epithelial structure could greatly improve early diagnosis and long-term monitoring in patients. Here, we have used tissue-engineered (TE) models of normal skin and oral mucosa to generate models of melanoma and oral cancer. We have used these to determine the ability of OCT to image epithelial differences in vitro. We report that while in vivo OCT gives reasonable depth information for both skin and oral mucosa, in vitro the information provided is less detailed but still useful. OCT can provide reassurance on the development of TE models of skin and oral mucosa as they develop in vitro. OCT was able to detect the gross alteration in the epithelium of skin and mucosal models generated with malignant cell lines but was less able to detect alteration in the epithelium of TE models that mimicked oral dysplasia or, in models where tumor cells had penetrated into the dermis.
Carcinoma, Squamous Cell/chemistry , Melanoma/chemistry , Mouth Mucosa/chemistry , Mouth Neoplasms/chemistry , Skin Neoplasms/chemistry , Tomography, Optical Coherence/methods , Cell Line, Tumor , Cells, Cultured , Fingers , Histocytochemistry , Humans , Models, Biological , Skin/chemistry
A near-infrared (NIR) absorbing contrast agent based on polypyrrole nanoparticles is described. Quantitative optical coherence tomography studies on tissue phantoms and Mie scattering calculations indicate their potential application for early-stage cancer diagnosis.
Contrast Media/pharmacology , Diagnostic Imaging/methods , Nanoparticles/chemistry , Neoplasms/diagnosis , Polymers/chemistry , Pyrroles/chemistry , Tomography, Optical Coherence/methods , Humans , Medical Oncology/methods , Models, Statistical , Phantoms, Imaging , Scattering, Radiation , Spectrophotometry, Ultraviolet/methods , Spectroscopy, Near-Infrared/methods
