We have been developing an automated method to image lymphatic vessels both ex vivo and in vivo with optical coherence tomography (OCT), using their optical transparency. Our method compensates for the OCT signal attenuation for each A-scan in combination with the correction of the confocal function and sensitivity fall-off, enabling reliable thresholding of lymphatic vessels from the OCT scans. Morphological image processing with a segment-joining algorithm is also incorporated into the method to mitigate partial-volume artifacts, which are particularly evident with small lymphatic vessels. Our method is demonstrated for two different clinical application goals: the monitoring of conjunctival lymphatics for surgical guidance and assessment of glaucoma treatment; and the longitudinal monitoring of human burn scars undergoing laser ablation treatment. We present examples of OCT lymphangiography ex vivo on porcine conjunctivas and in vivo on human burn scars, showing the visualization of the lymphatic vessel network and their longitudinal changes due to treatment.
Optical coherence elastography (OCE) is emerging as a potentially useful tool in the identification of a number of diseases. In our group, we are developing OCE techniques based on compressive loading. Typically, these techniques employ a quasi-static mechanical load introduced by uniaxially compressing a sample with a rigid plate. The resulting deformation of the sample is measured using phase-sensitive detection and the local axial strain is estimated from the slope of displacement over a finite depth in the sample, providing qualitative mechanical contrast. In this talk, an overview of our work will be given and some of the outstanding challenges described. Our group’s work in OCE can broadly be divided into four streams, each of which will be described in detail in the talk: system development; techniques; quantification; and applications.
• System development: The phase-sensitive OCE method we have developed will be described, as well as a high resolution optical coherence microscopy-based elastography system suitable for imaging cellular-scale mechanical properties.
• Techniques: In addition to presenting techniques to estimate strain, our approaches to imaging tissue viscoelasticity and nonlinearity will be described. A technique to segment elastograms based on strain heterogeneity will be presented.
• Quantification: Methods under development to quantify tissue stiffness in compression OCE will be described. This work is enabled by optical palpation and solutions to the forward and inverse elasticity problems.
• Applications: Three applications areas will be described: intraoperative assessment of tumour margins, mapping stiffness in tumour biology and assessing the stiffness of cardiovascular tissue in an animal model.
KEYWORDS: Tissues, Optical coherence tomography, Tissue optics, Birefringence, Tumors, In vivo imaging, Signal attenuation, Cancer, Microscopy, Medical imaging
INVITED TALK
Advances in imaging tissue microstructure in living subjects, or in freshly excised tissue with minimum preparation and processing, are important for future diagnosis and surgical guidance in the clinical setting, particularly for application to cancer. Whilst microscopy methods continue to advance on the cellular scale and medical imaging is well established on the scale of the whole tumor or organ, it is attractive to consider imaging the tumor environment on the micro-scale, between that of cells and whole tissues. Such a scenario is ideally suited to optical coherence tomography (OCT), with the twin attractions of requiring little or no tissue preparation, and in vivo capability. OCT’s intrinsic scattering contrast reveals many morphological features of tumors, but is frequently ineffective in revealing other important aspects, such as microvasculature, or in reliably distinguishing tumor from uninvolved stroma. To address these shortcomings, we are developing several advances on the basic OCT approach. We are exploring speckle fluctuations to image tissue microvasculature and we have been developing several parametric approaches to tissue micro-scale characterization. Our approaches extract, from a three-dimensional OCT data set, a two-dimensional image of an optical parameter, such as attenuation or birefringence, or a mechanical parameter, such as stiffness, that aids in characterizing the tissue. This latter method, termed optical coherence elastography, parallels developments in ultrasound and magnetic resonance imaging. Parametric imaging of birefringence and of stiffness both show promise in addressing the important issue of differentiating cancer from uninvolved stroma in breast tissue.
Probing the mechanical properties of skin at high resolution could aid in the assessment of skin pathologies by, for example, detecting the extent of cancerous skin lesions and assessing pathology in burn scars. Here, we present two elastography techniques based on optical coherence tomography (OCT) to probe the local mechanical properties of skin. The first technique, optical palpation, is a high-resolution tactile imaging technique, which uses a complaint silicone layer positioned on the tissue surface to measure spatially-resolved stress imparted by compressive loading. We assess the performance of optical palpation, using a handheld imaging probe on a skin-mimicking phantom, and demonstrate its use on human skin. The second technique is a strain imaging technique, phase-sensitive compression OCE that maps depth-resolved mechanical variations within skin. We show preliminary results of in vivo phase-sensitive compression OCE on a human skin lesion.
We demonstrate the first application of the recently proposed method of optical palpation to in vivo imaging of human skin. Optical palpation is a tactile imaging technique that probes the spatial variation of a sample’s mechanical properties by producing an en face map of stress measured at the sample surface. This map is determined from the thickness of a translucent, compliant stress sensor placed between a loading element and the sample and is measured using optical coherence tomography. We assess the performance of optical palpation using a handheld imaging probe on skin-mimicking phantoms, and demonstrate its use on human skin lesions. Our results demonstrate the capacity of optical palpation to delineate the boundaries of lesions and to map the mechanical contrast between lesions and the surrounding normal skin.
We demonstrate the in vivo assessment of human scars by parametric imaging of birefringence using polarization-sensitive optical coherence tomography (PS-OCT). Such in vivo assessment is subject to artifacts in the detected birefringence caused by scattering from blood vessels. To reduce these artifacts, we preprocessed the PS-OCT data using a vascular masking technique. The birefringence of the remaining tissue regions was then automatically quantified. Results from the scars and contralateral or adjacent normal skin of 13 patients show a correspondence of birefringence with scar type: the ratio of birefringence of hypertrophic scars to corresponding normal skin is 2.2±0.2 (mean±standard deviation), while the ratio of birefringence of normotrophic scars to normal skin is 1.1±0.4. This method represents a new clinically applicable means for objective, quantitative human scar assessment.
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