Quadruple ultrasound, photoacoustic, optical coherence, and fluorescence fusion imaging with a transparent ultrasound transducer.

Ultrasound and optical imagers are used widely in a variety of biological and medical applications. In particular, multimodal implementations combining light and sound have been actively investigated to improve imaging quality. However, the integration of optical sensors with opaque ultrasound transducers suffers from low signal-to-noise ratios, high complexity, and bulky form factors, significantly limiting its applications. Here, we demonstrate a quadruple fusion imaging system using a spherically focused transparent ultrasound transducer that enables seamless integration of ultrasound imaging with photoacoustic imaging, optical coherence tomography, and fluorescence imaging. As a first application, we comprehensively monitored multiparametric responses to chemical and suture injuries in rats' eyes in vivo, such as corneal neovascularization, structural changes, cataracts, and inflammation. As a second application, we successfully performed multimodal imaging of tumors in vivo, visualizing melanomas without using labels and visualizing 4T1 mammary carcinomas using PEGylated gold nanorods. We strongly believe that the seamlessly integrated multimodal system can be used not only in ophthalmology and oncology but also in other healthcare applications with broad impact and interest.

Stadium-type resonator sensor based on a multi-mode waveguide with mode discrimination phenomenon.

In this work, we present a multi-mode resonator based on SU-8 polymer and experimentally verify that the resonator showed mode discrimination can be used as a sensor with high performance. According to field emission scanning electron microscopy (FE-SEM) images, the fabricated resonator shows sidewall roughness which is canonically considered to be undesirable after a typical development process. In order to analyze the effect of sidewall roughness, we conduct the resonator simulation considering the roughness under various conditions. Mode discrimination still occurs even in the presence of sidewall roughness. In addition, waveguide width controllable by UV exposure time effectively contributes to mode discrimination. To verify the resonator as a sensor, we perform a temperature variation experiment, which results in a high sensitivity of about 630.8 nm/RIU. This result shows that the multi-mode resonator sensor fabricated via a simple process is competitive with other single-mode waveguide sensors.

Imaging Motion: A Comprehensive Review of Optical Coherence Tomography Angiography.

Optical coherence tomography (OCT) is a three-dimensional (3-D) optical imaging technology that provides noninvasive, micrometer resolution images of structural interiors within biological samples with an approximately 1 ~ 2 mm penetration depth. Over the last decades, advances in OCT have revolutionized biomedical imaging by demonstrating a potential of optical biopsy in preclinical and clinical settings. Recently, functional OCT imaging has shown a promise as angiography to visualize cell-perfused vasculatures in the tissue bed in vivo without requiring any exogenous contrast agents. This new technology termed OCT angiography (OCTA) possesses a unique imaging capability of delineating tissue morphology and blood or lymphatic vessels down to capillaries at real-time acquisition rates. For the past 10 years since 2007, OCTA has been proven to be a useful tool to identify disorder or dysfunction in tissue microcirculation from both experimental animal studies and clinical studies in ophthalmology and dermatology. In this section, we overview about OCTA including a basic principle of OCTA explained with simple optical physics, and its scan protocols and post-processing algorithms for acquisition of angiography. Then, potential and challenge of OCTA for clinical settings are shown with outcomes of human studies.

Multifunctional in vivo imaging for monitoring wound healing using swept-source polarization-sensitive optical coherence tomography.

BACKGROUND AND OBJECTIVE: Wound healing involves a complex and dynamic biological process in response to tissue injury. Monitoring of the cascade of cellular events is useful for wound management and treatment. The aim of this study is to demonstrate the potential of multifunctional polarization-sensitive optical coherence tomography (PS-OCT) to longitudinally monitor the self-healing process in a murine cutaneous wound model. MATERIALS AND METHODS: A multi-functional PS-OCT system based on swept source OCT configuration (1,310 nm central wavelength) was designed to obtain simultaneously microstructural, blood perfusion, and birefringent information of a biological tissue in vivo. A 1-mm-diameter wound was generated in a mouse pinna with a complete biopsy punch. Afterwards, the self-healing process of the injured tissue was observed every week over 6-week period using the multifunctional system to measure changes in the tissue birefringence. Further OCT angiography (OCTA) was used in post data processing to obtain blood perfusion information over the injured tissue. RESULTS: Three complementary images indicating the changes in anatomical, vascular, and birefringent information of tissue around wound were simultaneously provided from a 3-dimensional (3-D) PS-OCT data set during the wound repair over 1 month. Specifically, inflammatory and proliferative phases of wound healing were characterized by thickened epidermal tissue (from OCT images) and angiogenesis (from OCT angiography images) around wound. Also, it was observed that the regenerating tissues had highly realigned birefringent structures (from PS-OCT images). CONCLUSION: This preliminary study suggests that the proposed multi-functional imaging modality has a great potential to improve the understanding of wound healing through non-invasive, serial monitoring of vascular and tissue responses to injury. Lasers Surg. Med. 50:213-221, 2018. © 2017 Wiley Periodicals, Inc.

Microvascular imaging and monitoring of human oral cavity lesions in vivo by swept-source OCT-based angiography.

We report the development of optical coherence tomography- (OCT) based angiography (OCTA) to image blood flow within microcirculatory tissue beds in human oral cavity in vivo with a field of view at 10 mm × 10 mm. Three-dimensional (3D) structural and vascular images of labial mucosa tissue are obtained at a single 3D acquisition. Pathologic mucosal sites with mouth ulcers are examined using the OCT tomograms and angiograms, upon which to monitor the lesion healing process over a period of 2 weeks. Quantitative metrics of the capillary loop density within the lamina propria layer are evaluated, providing statistically significant difference between healthy and diseased conditions over time. Furthermore, tissue anatomy and vessel morphology of other susceptible sites to ulcer, such as tongue, alveolar mucosa, and labial frenulum, are also imaged to demonstrate the promise of the proposed method as a clinically useful tool for the diagnosis and monitoring of therapeutic treatment of oral tissue abnormalities.

Wide-field optical coherence tomography angiography enabled by two repeated measurements of B-scans.

Optical coherence tomography angiography (OCTA) has increasingly become clinically important, particularly in ophthalmology. However, the field of view (FOV) for current OCTA imaging is severely limited due to A-scan rates that can be afforded by current clinical systems and, more importantly, the requirement of a repeated scanning protocol. This Letter evaluates the possibility of using only two repeated B-scans for OCTA for the purpose of an increased FOV. The effect of repeated numbers on the OCTA result is discussed through experiments on an animal model in vivo and evaluated using quantitative metrics for image quality. Demonstrated through in vivo imaging of a pathological human eye, we show that optical microangiography-based OCTA with two repeated B-scans can provide wide-field angiography up to 12×12 mm with clinically acceptable image quality.

Potential use of OCT-based microangiography in clinical dermatology.

BACKGROUND: Optical coherence tomography (OCT) is a revolutionary imaging technique used commonly in ophthalmology, and on the way to become clinically viable alternative in dermatology due to its capability of acquiring histopathology level details of in vivo tissue, non-invasively. In this study, we demonstrate the capabilities of OCT-based microangiography in detecting high resolution, three-dimensional structural, and microvascular features of in vivo human skin with various conditions. METHODS: A swept-source OCT system that operates on a central wavelength of 1310 nm with an A-line rate of 100 kHz is used in this study. We apply optical microangiography (OMAG) technique to visualize the structural and microvascular changes in tissue. RESULTS: OMAG images provide detailed visualization of functional microvasculature of healthy human skin from cheek and forehead areas, abnormal skin conditions from face, chest and belly. Moreover, OMAG is capable of monitoring the progress of wound healing on human skin from arm, delivering unprecedented detail of microstructural and microvascular information during longitudinal wound healing process. CONCLUSION: The presented results promise the clinical use of OCT angiography, aiming to treat prevalent cutaneous diseases, by detecting blood perfusion, and structural changes within human skin, in vivo.

High resolution imaging of acne lesion development and scarring in human facial skin using OCT-based microangiography.

BACKGROUND AND OBJECTIVE: Acne is a common skin disease that often leads to scarring. Collagen and other tissue damage from the inflammation of acne give rise to permanent skin texture and microvascular changes. In this study, we demonstrate the capabilities of optical coherence tomography-based microangiography in detecting high-resolution, three-dimensional structural, and microvascular features of in vivo human facial skin during acne lesion initiation and scar development. MATERIALS AND METHODS: A real time swept source optical coherence tomography system is used in this study to acquire volumetric images of human skin. The system operates on a central wavelength of 1,310 nm with an A-line rate of 100 kHz, and with an extended imaging range (∼12 mm in air). The system uses a handheld imaging probe to image acne lesion on a facial skin of a volunteer. We utilize optical microangiography (OMAG) technique to evaluate the changes in microvasculature and tissue structure. RESULTS: Thanks to the high sensitivity of OMAG, we are able to image microvasculature up to capillary level and visualize the remodeled vessels around the acne lesion. Moreover, vascular density change derived from OMAG measurement is provided as an alternative biomarker for the assessment of human skin diseases. In contrast to other techniques like histology or microscopy, our technique made it possible to image 3D tissue structure and microvasculature up to 1.5 mm depth in vivo without the need of exogenous contrast agents. CONCLUSIONS: The presented results are promising to facilitate clinical trials aiming to treat acne lesion scarring, as well as other prevalent skin diseases, by detecting cutaneous blood flow and structural changes within human skin in vivo.

In vivo OCT microangiography of rodent iris.

We report on the functional optical coherence tomography (OCT) imaging of iris tissue morphology and microcirculation in living small animals. Anterior segments of healthy mouse and rat eyes are imaged with high-speed spectral domain OCT (SD-OCT) utilizing ultrahigh sensitive optical microangiography (UHS-OMAG) imaging protocol. 3D iris microvasculature is produced by the use of an algorithm that calculates absolute differences between the amplitudes of the OCT interframes. We demonstrate that the UHS-OMAG is capable of delineating iris microvascular beds in the mouse and rat with capillary-level resolution. Furthermore, the fast imaging speed enables dynamic imaging of iris micro-vascular response during drug-induced pupil dilation. We believe that this OCT angiographic approach has a great potential for in situ and in vivo monitoring of the microcirculation within iris tissue beds in rodent disease models that have microvascular involvement.

Analysis of cross-sectional image filters for evaluating nonaveraged optical microangiography images.

Optical microangiography (OMAG) is a method that enables the noninvasive extraction of blood vessels within biological tissues. OMAG B-frames are prone to noise; therefore, techniques such as B-frame averaging have been applied to reduce these effects. A drawback of this method is that the total acquisition time and amount of data collected are increased; hence, the data are susceptible to motion artifacts and decorrelation. In this paper we propose using an image filter on a nonaveraged OMAG B-frame to reduce its noise. Consequently, B-frames comparable to the averaged OMAG B-frame are obtained, while reducing the total acquisition and processing time. The method is tested with two different systems, a high-resolution spectral domain and a relatively low-resolution swept-source optical coherence tomography system. It is demonstrated that the weighted average filter produces the lowest B-frame error; however, all filters produce comparable results when quantifying the en face projection view image.

Enhancing the color gamut of waveguide displays for augmented reality head-mounted displays through spatially modulated diffraction grating.

Augmented reality (AR) applications require displays with an extended color gamut to facilitate the presentation of increasingly immersive content. The waveguide (WG) display technology, which is typical AR demonstration method, is a critical constraint on the color gamut of AR systems because of the intrinsic properties of the holographic optical elements (HOEs) used in this technology. To overcome this limitation, we introduce a method of spatially modulated diffractive optics that can expand the color gamut of HOE-based WG displays. This approach involves spatial modulation using sub-pixelized HOEs, which enables the diffraction of red, green, and blue rays along identical directions. The proposed structure considers both the characteristics of the HOE and the wavelength sensitivity of the observer to optimize the color gamut. Consequently, an expanded color gamut was achieved. The results of the theoretical and experimental analyses substantiate the effectiveness and practicality of this method in enhancing the color gamut of HOE-based WG displays. Thus, the proposed method can facilitate the implementation of more immersive AR displays.

Refractive index measurements of multiple layers using numerical refocusing in FF-OCT.

We propose and demonstrate the novel method of refractive index (RI) measurement for each layer of multilayered samples, which is based on numerical refocusing in full field optical coherence tomography (FF-OCT). The en-face FF-OCT image on an inner layer boundary of a multilayered sample is unintentionally blurred or defocused due to the RI of the sample itself, but can be numerically refocused. The refocusing is performed by numerically shifting the image sensor plane of the system, in general. However, by calculating the corresponding sample shift and then compared it with the actual sample shifting distance, we could extract the average RI of the layer between any two layer boundaries within the multilayered sample. In addition, the thickness of that particular layer could be derived at the same time. For the idea proof, several samples were prepared by stacking, for each sample, two transparent plates with a gap in between. While changing the material of the plate and filling the gap with oil, the RIs of the plate and the oil were measured. For oils of various RIs, from 1.2977 to 1.3857, the measured RIs were well matched with the reported ones within 0.205%. Moreover, even with a stack of various and multiple plates in front of the same oil layer, the oil RI and the physical thickness of the oil layer were extracted with average errors of only 0.065% and 0.990%, respectively.

Fast mapping of absorbing defects in optical materials by full-field photothermal reflectance microscopy.

We report a technique for rapidly mapping absorbing defects in optical materials, which act as laser-induced damage precursors, based on full-field photothermal reflectance microscopy. An intensity-modulated pump beam heats absorbing defects in the optical sample, creating localized, modulated refractive-index variations around the defects. A probe beam then illuminates the defect sites, and the measured amplitude of the reflectance variation is used to map the distribution of defects in the medium. Measurements show that this method offers a faster defect mapping speed of about 0.03 mm(2) per minute and a detectivity of a few tens of nanometers comparable to that of conventional scanning photothermal deflection microscopy.

High-speed thermoreflectance microscopy using charge-coupled device-based Fourier-domain filtering.

We present a Fourier-domain filtering method for charge-coupled device (CCD)-based thermoreflectance microscopy to improve the thermal imaging speed while maintaining high thermal sensitivity. The time-varying reflected light distribution from the surface of bias-modulated microresistor was recorded by a CCD camera in free-run mode and converted to the frequency domain using the fast Fourier transform (FFT) for all pixels of the CCD. After frequency peak filtering followed by inverse FFT, a thermoreflectance image was obtained. The imaging results of the proposed method were quantitatively compared with those of the conventional four-bucket method, showing that the Fourier-domain filtering method can provide thermal imaging 24-42 times faster than the four-bucket method, depending on the required thermal sensitivity.

Non-Invasive Monitoring of Cutaneous Wound Healing in Non-Diabetic and Diabetic Model of Adult Zebrafish Using OCT Angiography.

A diabetic wound presents a severe risk of infections and other complications because of its slow healing. Evaluating the pathophysiology during wound healing is imperative for wound care, necessitating a proper diabetic wound model and assay for monitoring. The adult zebrafish is a rapid and robust model for studying human cutaneous wound healing because of its fecundity and high similarities to human wound repair. OCTA as an assay can provide three-dimensional (3D) imaging of the tissue structure and vasculature in the epidermis, enabling monitoring of the pathophysiologic alterations in the zebrafish skin wound. We present a longitudinal study for assessing the cutaneous wound healing of the diabetic adult zebrafish model using OCTA, which is of importance for the diabetes research using the alternative animal models. We used non-diabetic (n = 9) and type 1 diabetes mellitus (DM) adult zebrafish models (n = 9). The full-thickness wound was generated on the fish skin, and the wound healing was monitored with OCTA for 15 days. The OCTA results demonstrated significant differences between diabetic and non-diabetic wound healing, involving delayed tissue remodeling and impaired angiogenesis for the diabetic wound, leading to slow wound recovery. The adult zebrafish model and OCTA technique may benefit long-term metabolic disease studies using zebrafish for drug development.

Smartphone-Based Rigid Endoscopy Device with Hemodynamic Response Imaging and Laser Speckle Contrast Imaging.

Modern smartphones have been employed as key elements in point-of-care (POC) devices due to remarkable advances in their form factor, computing, and display performances. Recently, we reported a combination of the smartphone with a handheld endoscope using laser speckle contrast imaging (LSCI), suggesting potential for functional POC endoscopy. Here, we extended our work to develop a smartphone-combined multifunctional handheld endoscope using dual-wavelength LSCI. Dual-wavelength LSCI is used to monitor the changes in dynamic blood flow as well as changes in the concentration of oxygenated (HbO2), deoxygenated (Hbr), and total hemoglobin (HbT). The smartphone in the device performs fast acquisition and computation of the raw LSCI data to map the blood perfusion parameters. The flow imaging performance of the proposed device was tested with a tissue-like flow phantom, exhibiting a speckle flow index map representing the blood perfusion. Furthermore, the device was employed to assess the blood perfusion status from an exteriorized intestine model of rat in vivo during and after local ischemia, showing that blood flow and HbO2 gradually decreased in the ischemic region whereas hyperemia and excess increases in HbO2 were observed in the same region right after reperfusion. The results indicate that the combination of LSCI with smartphone endoscopy delivers a valuable platform for better understanding of the functional hemodynamic changes in the vasculatures of the internal organs, which may benefit POC testing for diagnosis and treatment of vascular diseases.

Health Status and Activity Discomfort among Elderly Drivers: Reality of Health Awareness.

As the number of elderly drivers rapidly increases worldwide, interest in the dangers of driving is growing as accidents rise. The purpose of this study was to conduct a statistical analysis of the driving risk factors of elderly drivers. In this analysis, data from the government organization's open data were used for the secondary processing of 10,097 people. Of the 9990 respondents, 2168 were current drivers, 1552 were past drivers but were not driving presently, and 6270 did not have a driver's license; the participants were divided into groups accordingly. The elderly drivers who were current drivers had a better subjective health status than those who were not. Visual and hearing aids were used in the current driving group, and their depression symptoms reduced as they drove. The elderly who were current drivers experienced difficulties while driving in terms of decreased vision, hearing loss, reduced arm/leg reaction speed, decreased judgment of the road conditions such as signals and intersections, and a decreased sense of speed. The results suggest that elderly drivers are unaware of the medical conditions that can negatively affect their driving. This study contributes to the safety management of elderly drivers by understanding their mental and physical status.

Tomographic imaging of a suspending single live cell using optical tweezer-combined full-field optical coherence tomography.

We propose a label-free depth-resolved tomographic scheme for imaging a single live cell in fluid. This approach utilizes a modified time-domain full-field optical coherence tomography (FF-OCT) system combined with an optical tweezer technique. The optical trap for holding a moving specimen is made by tightly focusing a 1064 nm Q-switching pulsed laser beam with a 1.0 NA microscope objective in the sample arm of the FF-OCT part. By cosharing the probe for both systems, the optical actions of trapping and cellular resolution tomographic imaging could be achieved simultaneously. Feasibility of the combined system is demonstrated by imaging micron-sized polystyrene beads and a living suspension cell in medium.

Automated counting of cerebral penetrating vessels using optical coherence tomography images of a mouse brain in vivo.

RATIONALE AND OBJECTIVES: Penetrating blood vessels emanating from cortical surface vasculature and lying deep in the cortex are essential vascular conduits for the shuttling of blood from superficial pial vessels to the capillary beds in parenchyma for the nourishment of neuronal brain tissues. Locating and counting the penetrating vessels is beneficial for the quantification of a course of ischemia in blood occlusive events such as stroke. This paper seeks to demonstrate and validate a method for automated penetrating vessel counting that uses optical coherence tomography (OCT). MATERIALS AND METHODS: This paper proposes an OCT method that effectively identifies and grades the cortical penetrating vessels in perfusion. The key to the proposed method is the harnessing of vascular features found in the penetrating vessels, which are distinctive from those of other vessels. In particular, with an increase in the light attenuation and flow turbulence, the contrast in the mean projection of the OCT datacube decreases, whereas that in the maximum projection of the Doppler frequency variance datacube increases. By multiplying the inversion of the former with the latter, its binary thresholding is sufficient to highlight the penetrating vessels and allows for their counting over the projection image. RESULTS: A computational method that leverages the decrease in mean OCT projection intensity and the increase in Doppler frequency variance at the penetrating vessel is developed. It successfully identifies and counts penetrating vessels with a high accuracy of over 87%. The penetrating vessel density is observed to be significantly reduced in the mouse model of focal ischemic stroke. CONCLUSION: The OCT analysis is effective for counting penetrating blood vessels in mice brains and may be applied to the rapid diagnosis and treatment of stroke in stroke models of small animals.

Compact Smartphone-Based Laser Speckle Contrast Imaging Endoscope Device for Point-of-Care Blood Flow Monitoring.

Laser speckle contrast imaging (LSCI) is a powerful visualization tool for quantifying blood flow in tissues, providing simplicity of configuration, ease of use, and intuitive results. With recent advancements, smartphone and camera technologies are suitable for the development of smartphone-based LSCI applications for point-of-care (POC) diagnosis. A smartphone-based portable LSCI endoscope system was validated for POC diagnosis of vascular disorders. The endoscope consisted of compact LED and laser illumination, imaging optics, and a flexible fiberscope assembled in a 3D-printed hand-held cartridge for access to body cavities and organs. A smartphone's rear camera was mounted thereto, enabling endoscopy, LSCI image acquisition, and processing. Blood flow imaging was calibrated in a perfused tissue phantom consisting of a microparticle solution pumped at known rates through tissue-mimicking gel and validated in a live rat model of BBN-induced bladder cancer. Raw LSCI images successfully visualized phantom flow: speckle flow index showed linearity with the pump flow rate. In the rat model, healthy and cancerous bladders were distinguishable in structure and vasculature. The smartphone-based low-cost portable mobile endoscope for monitoring blood flow and perfusion shows promise for preclinical applications and may be suitable for primary diagnosis at home or as a cost-effective POC testing assay.