A Dorsal Skinfold Window Chamber Tumor Mouse Model for Combined Intravital Microscopy and Magnetic Resonance Imaging in Translational Cancer Research.

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the tumor microenvironment and its response to therapy. These imaging modalities have micron-scale resolution but have limited use in the clinic due to their shallow penetration depth into tissue. More clinically applicable imaging modalities such as CT, MRI, and PET have much greater penetration depth but have comparatively lower spatial resolution (mm scale). To translate preclinical intravital imaging findings into the clinic, new methods must be developed to bridge this micro-to-macro resolution gap. Here we describe a dorsal skinfold window chamber tumor mouse model designed to enable preclinical intravital and clinically applicable (CT and MR) imaging in the same animal, and the image analysis platform that links these two disparate visualization methods. Importantly, the described window chamber approach enables the different imaging modalities to be co-registered in 3D using fiducial markers on the window chamber for direct spatial concordance. This model can be used for validation of existing clinical imaging methods, as well as for the development of new ones through direct correlation with "ground truth" high-resolution intravital findings. Finally, the tumor response to various treatments-chemotherapy, radiotherapy, photodynamic therapy-can be monitored longitudinally with this methodology using preclinical and clinically applicable imaging modalities. The dorsal skinfold window chamber tumor mouse model and imaging platforms described here can thus be used in a variety of cancer research studies, for example, in translating preclinical intravital microscopy findings to more clinically applicable imaging modalities such as CT or MRI.

Maximizing atmospheric-disturbed fiber coupling efficiency with speckle-based phase retrieval and a single-pixel camera.

An approach to adaptive optics utilizing a single-pixel camera (SPC) is proposed to maximize fiber coupling efficiency at the receiver side of an optical satellite-to-ground link perturbed by atmospheric turbulence. Using a single-pixel wavefront sensor enables operation at longer optical wavelengths, such as near and far infrared, which have advantageous propagation characteristics for free space optical communication. In this approach, a focal plane intensity image of the atmospheric-disturbed wavefront is taken via an SPC using a compressed sensing technique. An iterative speckle-based phase retrieval algorithm is then applied to infer the phase distortion corrected by a deformable mirror in a feedback loop. This computational approach to inferring the phase of the wavefront overcomes the limitations of traditional Shack-Hartman-based approaches, which are difficult to implement at high speed and at the long infrared wavelengths proposed for future optical satellite communication downlinks. It has been shown that fiber coupling efficiency is increased from less than 5% to 40%-50% in medium-to-strong turbulence scenarios with the phase retrieval algorithm proposed in this work.

Binary dose level classification of tumour microvascular response to radiotherapy using artificial intelligence analysis of optical coherence tomography images.

The dominant consequence of irradiating biological systems is cellular damage, yet microvascular damage begins to assume an increasingly important role as the radiation dose levels increase. This is currently becoming more relevant in radiation medicine with its pivot towards higher-dose-per-fraction/fewer fractions treatment paradigm (e.g., stereotactic body radiotherapy (SBRT)). We have thus developed a 3D preclinical imaging platform based on speckle-variance optical coherence tomography (svOCT) for longitudinal monitoring of tumour microvascular radiation responses in vivo. Here we present an artificial intelligence (AI) approach to analyze the resultant microvascular data. In this initial study, we show that AI can successfully classify SBRT-relevant clinical radiation dose levels at multiple timepoints (t = 2-4 weeks) following irradiation (10 Gy and 30 Gy cohorts) based on induced changes in the detected microvascular networks. Practicality of the obtained results, challenges associated with modest number of animals, their successful mitigation via augmented data approaches, and advantages of using 3D deep learning methodologies, are discussed. Extension of this encouraging initial study to longitudinal AI-based time-series analysis for treatment outcome predictions at finer dose level gradations is envisioned.

Longitudinal in-vivo quantification of tumour microvascular heterogeneity by optical coherence angiography in pre-clinical radiation therapy.

Stereotactic body radiotherapy (SBRT) is an emerging cancer treatment due to its logistical and potential therapeutic benefits as compared to conventional radiotherapy. However, its mechanism of action is yet to be fully understood, likely involving the ablation of tumour microvasculature by higher doses per fraction used in SBRT. In this study, we hypothesized that longitudinal imaging and quantification of the vascular architecture may elucidate the relationship between the microvasculature and tumour response kinetics. Pancreatic human tumour xenografts were thus irradiated with single doses of [Formula: see text], [Formula: see text] and [Formula: see text] Gy to simulate the first fraction of a SBRT protocol. Tumour microvascular changes were monitored with optical coherence angiography for up to [Formula: see text] weeks following irradiation. The temporal kinetics of two microvascular architectural metrics were studied as a function of time and dose: the diffusion-limited fraction, representing poorly vascularized tissue [Formula: see text] µm from the nearest detected vessel, and the vascular distribution convexity index, a measure of vessel aggregation at short distances. These biological metrics allowed for dose dependent temporal evaluation of tissue (re)vascularization and vessel aggregation after radiotherapy, showing promise for determining the SBRT dose-response relationship.

Bridging the macro to micro resolution gap with angiographic optical coherence tomography and dynamic contrast enhanced MRI.

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) is emerging as a valuable tool for non-invasive volumetric monitoring of the tumor vascular status and its therapeutic response. However, clinical utility of DCE-MRI is challenged by uncertainty in its ability to quantify the tumor microvasculature ([Formula: see text] scale) given its relatively poor spatial resolution (mm scale at best). To address this challenge, we directly compared DCE-MRI parameter maps with co-registered micron-scale-resolution speckle variance optical coherence tomography (svOCT) microvascular images in a window chamber tumor mouse model. Both semi and fully quantitative (Toft's model) DCE-MRI metrics were tested for correlation with microvascular svOCT biomarkers. svOCT's derived vascular volume fraction (VVF) and the mean distance to nearest vessel ([Formula: see text]) metrics were correlated with DCE-MRI vascular biomarkers such as time to peak contrast enhancement ([Formula: see text] and [Formula: see text] respectively, [Formula: see text] for both), the area under the gadolinium-time concentration curve ([Formula: see text] and [Formula: see text] respectively, [Formula: see text] for both) and [Formula: see text] ([Formula: see text] and [Formula: see text] respectively, [Formula: see text] for both). Several other correlated micro-macro vascular metric pairs were also noted. The microvascular insights afforded by svOCT may help improve the clinical utility of DCE-MRI for tissue functional status assessment and therapeutic response monitoring applications.

Volumetric tumor delineation and assessment of its early response to radiotherapy with optical coherence tomography.

Texture analyses of optical coherence tomography (OCT) images have shown initial promise for differentiation of normal and tumor tissues. This work develops a fully automatic volumetric tumor delineation technique employing quantitative OCT image speckle analysis based on Gamma distribution fits. We test its performance in-vivo using immunodeficient mice with dorsal skin window chambers and subcutaneously grown tumor models. Tumor boundaries detection is confirmed using epi-fluorescence microscopy, combined photoacoustic-ultrasound imaging, and histology. Pilot animal study of tumor response to radiotherapy demonstrates high accuracy, objective nature, novelty of the proposed method in the volumetric separation of tumor and normal tissues, and the sensitivity of the fitting parameters to radiation-induced tissue changes. Overall, the developed methodology enables hitherto impossible longitudinal studies for detecting subtle tissue alterations stemming from therapeutic insult.

Analysis of low-scattering regions in optical coherence tomography: applications to neurography and lymphangiography.

Analysis of semi-transparent low scattering biological structures in optical coherence tomography (OCT) has been actively pursued in the context of lymphatic imaging, with most approaches relying on the relative absence of signal as a means of detection. Here we present an alternate methodology based on spatial speckle statistics, utilizing the similarity of a distribution of given voxel intensities to the power distribution function of pure noise, to visualize the low-scattering biological structures of interest. In a human tumor xenograft murine model, we show that these correspond to lymphatic vessels and nerves; extensive histopathologic validation studies are reported to unequivocally establish this correspondence. The emerging possibility of OCT lymphangiography and neurography is novel and potentially impactful (especially the latter), although further methodology refinement is needed to distinguish between the visualized lymphatics and nerves.

Preclinical quantitative in-vivo assessment of skin tissue vascularity in radiation-induced fibrosis with optical coherence tomography.

Radiation therapy (RT) is widely and effectively used for cancer treatment but can also cause deleterious side effects, such as a late-toxicity complication called radiation-induced fibrosis (RIF). Accurate diagnosis of RIF requires analysis of histological sections to assess extracellular matrix infiltration. This is invasive, prone to sampling limitations, and thus rarely used; instead, current practice relies on subjective clinical surrogates, including visual observation, palpation, and patient symptomatology questionnaires. This preclinical study demonstrates that functional optical coherence tomography (OCT) is a useful tool for objective noninvasive in-vivo assessment and quantification of fibrosis-associated microvascular changes in tissue. Data were collected from murine hind limbs 6 months after 40-Gy single-dose irradiation and compared with nonirradiated contralateral tissues of the same animals. OCT-derived vascular density and average vessel diameter metrics were compared to quantitative vascular analysis of stained histological slides. Results indicate that RIF manifests significant microvascular changes at this time point posttreatment. Abnormal microvascular changes visualized by OCT in this preclinical setting suggest the potential of this label-free high-resolution noninvasive functional imaging methodology for RIF diagnosis and assessment in the context of clinical RT.

Preclinical longitudinal imaging of tumor microvascular radiobiological response with functional optical coherence tomography.

Radiation therapy (RT) is widely used for cancer treatment, alone or in combination with other therapies. Recent RT advances have revived interest in delivering higher dose in fewer fractions, which may invoke both cellular and microvascular damage mechanisms. Microvasculature may thus be a potentially sensitive functional biomarker of RT early response, especially for such emerging RT treatments. However it is difficult to measure directly and non-invasively, and its time course, dose dependencies, and overall importance in tumor control are unclear. We use functional optical coherence tomography for quantitative longitudinal in vivo imaging in preclinical models of human tumor xenografts subjected to 10, 20 and 30 Gy doses, furnishing a detailed assessment of vascular remodeling following RT. Immediate (minutes to tens of minutes) and early (days to weeks) RT responses of microvascular supply, as well as tumor volume and fluorescence intensity, were quantified and demonstrated robust and complex temporal dose-dependent behaviors. The findings were compared to theoretical models proposed in the literature.

Texture analysis of optical coherence tomography speckle for characterizing biological tissues in vivo.

We demonstrate a method for differentiating tissue disease states using the intrinsic texture properties of speckle in optical coherence tomography (OCT) images of normal and tumor tissues obtained in vivo. This approach fits a gamma distribution function to the nonlog-compressed OCT image intensities, thus allowing differentiation of normal and tumor tissues in an ME-180 human cervical cancer mouse xenograft model. Quantitative speckle intensity distribution analysis thus shows promise for identifying tissue pathologies, with potential for early cancer detection in vivo.

Stationary-fiber rotary probe with unobstructed 360° view for optical coherence tomography.

A side-scanning fiber probe is a critical component for optical coherence tomography in medical imaging and diagnosis. We propose and fabricate an on-axis rotating probe that performs in situ, circumferential scanning that is shadow-free (not susceptible to shadow effects caused by the motor's wires). A miniature motor that incorporates a bored-out shaft for the optical fiber is located at the distal end of the probe, which results in a more stable and uniform circumferential scan, free from wire-shadow interference effects. More importantly, this design, novel to our knowledge, compared to other probes avoids the insertion losses introduced by optical coupling components and the multitude of optical interfaces, which is very important for sensing weak signals backscattered from structures deep in the tissue.

Simultaneous dual-wavelength-band common-path swept-source optical coherence tomography with single polygon mirror scanner.

We report a novel (to the best of our knowledge) simultaneous 1310/1550 two-wavelength band swept laser source and dual-band common-path swept-source optical coherence tomography (SS-OCT). Synchronized dual-wavelength tuning is performed by using two laser cavities and narrowband wavelength filters with a single dual-window polygonal scanner. Measured average output powers of 60 and 27 mW have been achieved for the 1310 and 1550 nm bands, respectively, while the two wavelengths were swept simultaneously from 1227 to 1387 nm for the 1310 nm band and from 1519 to 1581 nm for the 1550 nm band at an A-scan rate of 65 kHz. Broadband wavelength-division multiplexing is used for coupling two wavelengths into a common-path single-mode GRIN-lensed fiber probe to form dual-band common-path SS-OCT. Simultaneous OCT imaging at 1310 and 1550 nm is achieved. This technique allows for in vivo high-speed OCT imaging with potential application in functional (spectroscopic) investigations.

Optical coherence tomography: fundamental principles, instrumental designs and biomedical applications.

The advances made in the last two decades in interference technologies, optical instrumentation, catheter technology, optical detectors, speed of data acquisition and processing as well as light sources have facilitated the transformation of optical coherence tomography from an optical method used mainly in research laboratories into a valuable tool applied in various areas of medicine and health sciences. This review paper highlights the place occupied by optical coherence tomography in relation to other imaging methods that are used in medical and life science areas such as ophthalmology, cardiology, dentistry and gastrointestinal endoscopy. Together with the basic principles that lay behind the imaging method itself, this review provides a summary of the functional differences between time-domain, spectral-domain and full-field optical coherence tomography, a presentation of specific methods for processing the data acquired by these systems, an introduction to the noise sources that plague the detected signal and the progress made in optical coherence tomography catheter technology over the last decade.

Signal attenuation and box-counting fractal analysis of optical coherence tomography images of arterial tissue.

The sensitivity of optical coherence tomography images to sample morphology is tested by two methods. The first method estimates the attenuation of the OCT signal from various regions of the probed tissue. The second method uses a box-counting algorithm to calculate the fractal dimensions in the regions of interest identified in the images. Although both the attenuation coefficient as well as the fractal dimension correlate very well with the anatomical features of the probed samples; the attenuation method provides a better sensitivity. Two types of samples are used in this study: segments of arteries collected from atherosclerosis-prone Watanabe rabbits (WHHL-MI) and healthy segments of porcine coronary arteries.

3x3 Mach-Zehnder interferometer with unbalanced differential detection for full-range swept-source optical coherence tomography.

Quadrature interferometry based on 3x3 fiber couplers could be used to double the effective imaging depth in swept-source optical coherence tomography. This is due to its ability to suppress the complex conjugate artifact naturally. We present theoretical and experimental results for a 3x3 Mach-Zehnder interferometer using a new unbalanced differential optical detection method. The new interferometer provides simultaneous access to complementary phase components of the complex interferometric signal. No calculations by trigonometric relationships are needed. We demonstrate a complex conjugate artifact suppression of 27 dB obtained in swept-source optical coherence tomography using our unbalanced differential detection. We show that our unbalanced differential detection has increased the signal-to-noise ratio by at least 4 dB compared to the commonly used balanced detection technique. This is due to better utilization of optical power.

Statistics of the depth-scan photocurrent in time-domain optical coherence tomography.

We derive the time-variant second-order statistics of the depth-scan photocurrent in time-domain optical coherence tomography (TD-OCT) systems using polarized thermal light sources and superluminescent diodes (SLDs). Since the asymptotic-joint-probability-distribution function (JPDF) of the photocurrent due to polarized thermal light is Gaussian and the signal-noise-ratio in TD-OCT is typically high (>80 dB), the JPDF of the depth-scan photocurrent could be approximated as a Gaussian random process that is completely determined by its second-order statistics. We analyze both direct and differential light detection schemes and include the effect of electronic thermal fluctuations. Our results are a necessary prerequisite for future development of statistical image processing techniques for TD-OCT.

Graded-index fiber lens proposed for ultrasmall probes used in biomedical imaging.

The quality and parameters of probing optical beams are extremely important in biomedical imaging systems both for image quality and light coupling efficiency considerations. For example, the shape, size, focal position, and focal range of such beams could have a great impact on the lateral resolution, penetration depth, and signal-to-noise ratio of the image in optical coherence tomography. We present a beam profile characterization of different variations of graded-index (GRIN) fiber lenses, which were recently proposed for biomedical imaging probes. Those GRIN lens modules are made of a single mode fiber and a GRIN fiber lens with or without a fiber spacer between them. We discuss theoretical analysis methods, fabrication techniques, and measured performance compared with theory.

Image enhancement for multilayer information retrieval by using full-field optical coherence tomography.

When a full-field optical coherence tomography (OCT) system is used to extract tomographic images from a multilayer information carrier, the resulting images may suffer from interlayer modulations and parasitic patterns derived from interference fringes. We describe and analyze these negative influences that degrade the quality of extracted tomographic images and propose practical algorithms and methods to minimize them. The emphasis of the discussion will be the removal of the parasitic fringes produced by the imperfection of a CCD camera. The simulative and experimental results of image enhancement for multilayer tomography extraction using full-field OCT are provided.

Relative measurements of second-order susceptibility with reflective second-harmonic generation.

There is a strong demand for a simple and reliable technique for second-order susceptibility measurements of thin films. Since the Maker fringe technique is limited to transparent substrates we propose an experimental protocol based on reflective second-harmonic generation (SHG). The proposed protocol is based on relative measurements of Z-cut quartz. An analytical expression of the reflective SHG intensity dependence of the polarizer, analyzer, and sample azimuth is presented. An error analysis is also presented. Thin organic film of the side-chain polymer poly(Disperse Red 1 Methacrylate-Co-Methyl-Methacrylate) is investigated. Results for different wavelengths are reported.