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Background and objective Spectrally encoded endoscopy (SEE) is an optical imaging technology that uses spatial wavelength multiplexing to conduct endoscopy in miniature, small diameter probes. Contrary to the previous side-viewing SEE devices, forward-viewing SEE probes are advantageous as they provide a look ahead that facilitates navigation and surveillance. The objective of this work was to develop a miniature forward-viewing SEE probe with a wide field of view and a high spatial resolution. Materials and methods We designed and developed a forward-viewing SEE device with an overall total diameter of 1.27 mm, which consists of a monolithic illumination probe with a length of 3.87 mm and a diameter of 500 µm, 8 multimode detection fibers that were polished at a 17° angle, a rotational scanning mechanism, and a sheath. The SEE device was evaluated using a USAF resolution target and was used for preclinical imaging of a swine joint ex vivo. Results This design resulted in a high resolution probe (best spatial resolution of 20.3 µm), a wide total angular field of view of 100°, and an effective number of imaging elements of ~344,000 pixels. The SEE probe performance was compared to a commercial color chip-on-the-tip endoscope; while monochrome, results showed better spatial resolution and a wider field of view for the SEE device. Conclusion These results demonstrate the potential of this forward-viewing SEE probe for visualization and navigation in medical imaging applications. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Optical microscopy has been widely used in biomedical research as it provides photophysical and photochemical information of the target in subcellular spatial resolution without requiring physical contact with the specimen. To obtain a deeper understanding of biological phenomena, several efforts have been expended to combine such optical imaging modalities into a single microscope system. However, the use of multiple light sources and detectors through separated beam paths renders previous systems extremely complicated or slow for in vivo imaging. Herein, we propose a novel high-speed multimodal optical microscope system that simultaneously visualizes five different microscopic contrasts, i.e., two-photon excitation, second-harmonic generation, backscattered light, near-infrared fluorescence, and fluorescence lifetime, using a single femtosecond pulsed laser. Our proposed system can visualize five modal images with a frame rate of 3.7 fps in real-time, thereby providing complementary optical information that enhances both structural and functional contrasts. This highly photon-efficient multimodal microscope system enables various properties of biological tissues to be assessed.

Spectrally encoded endoscopy (SEE) is an ultra-miniature endoscopy technology that encodes each spatial location on the sample with a different wavelength. One challenge in SEE is achieving color imaging with a small probe. We present a novel SEE probe that is capable of conducting real-time RGB imaging using three diffraction orders (6th order diffraction of the blue spectrum, 5th of green, and 4th of red). The probe was comprised of rotating 0.5 mm-diameter illumination optics inside a static, 1.2 mm-diameter flexible sheath with a rigid distal length of 5 mm containing detection fibers. A color chart, resolution target, and swine tissue were imaged. The device achieved 44k/59k/23k effective pixels per R/G/B channels over a 58° angular field and differentiated a wide gamut of colors.

Rationale: Atherosclerotic plaque is a chronic inflammatory disorder involving lipid accumulation within arterial walls. In particular, macrophages mediate plaque progression and rupture. While PPARγ agonist is known to have favorable pleiotropic effects on atherogenesis, its clinical application has been very limited due to undesirable systemic effects. We hypothesized that the specific delivery of a PPARγ agonist to inflamed plaques could reduce plaque burden and inflammation without systemic adverse effects. Methods: Herein, we newly developed a macrophage mannose receptor (MMR)-targeted biocompatible nanocarrier loaded with lobeglitazone (MMR-Lobe), which is able to specifically activate PPARγ pathways within inflamed high-risk plaques, and investigated its anti-atherogenic and anti-inflammatory effects both in in vitro and in vivo experiments. Results: MMR-Lobe had a high affinity to macrophage foam cells, and it could efficiently promote cholesterol efflux via LXRα-, ABCA1, and ABCG1 dependent pathways, and inhibit plaque protease expression. Using in vivo serial optical imaging of carotid artery, MMR-Lobe markedly reduced both plaque burden and inflammation in atherogenic mice without undesirable systemic effects. Comprehensive analysis of en face aorta by ex vivo imaging and immunostaining well corroborated the in vivo findings. Conclusion: MMR-Lobe was able to activate PPARγ pathways within high-risk plaques and effectively reduce both plaque burden and inflammation. This novel targetable PPARγ activation in macrophages could be a promising therapeutic strategy for high-risk plaques.

Coronary arteries are covered by a layer of endothelial cells (ECs) that have a thickness of approximately 1 μm. Impairment of ECs is at the origin of coronary atherosclerosis and its clinical manifestations. The current gold standard, scanning electron microscopy (SEM), has demonstrated that ECs

This paper presents a novel XY nano-positioning stage using a compliant parallel mechanism with small crosstalk and yaw motion. A parallel mechanism can implement multi-degree-of-freedom motion with a small footprint. However, this implies crosstalk between the motion axes. In an effort to reduce crosstalk and yaw motion, we proposed a novel flexure guide structure. The proposed stage consists of two flexure displacement amplifiers driven by piezoelectric actuators, two combined double four-bar flexure guides, and two additional parallelogram guides. To prevent the rotational deformation of one amplifier due to the transverse force of the other, we introduce an intermediate leaf spring at the output ends of the amplifiers. This new parallel mechanism is optimally designed based on an analytical model of the stage to assure small crosstalk and yaw motion. The analytical model is verified as being well-described using a finite element analysis. An XY nano-positioning stage of size 150 × 150 × 30 mm3 is fabricated following the optimal design. Experiments are carried out to verify the static and dynamic performances of the proposed XY nano-positioning stage. The proposed stage has an X and Y directional motion range of 120 μm in closed loop. The resolution of the stage is 3 nm in both the X and Y directional motions. The actual crosstalk and yaw were limited to 0.770 μm and 13.5 μrad, while in the optimal design they were estimated at 0.5 μm and 15 μrad, respectively.

The use of adhesive water-in-oil (W/O) emulsions covered with phospholipids have been limited due to their poor stability. We suggest a new and simple method to create adhesive W/O emulsions stabilized by 1,2-dioleoyl- sn -glycero-3-phosphocholine (DOPC) with two different cosurfactants (docosahexaenoic acid (DHA) and sorbitan oleate (SPAN 80)). Although an adhesive W/O emulsion with DOPC is typically unstable because of its molecular structure, we demonstrate that the addition of cosurfactants whose molecular shapes could be complementary to that of DOPC far better stabilizes W/O emulsions and indeed leads to the production of adhesive emulsions by the formation of a bilayer between two monolayers of each droplet surface.

The magnetically levitated system, so called maglev system, has been researched and developed with the purpose of vacuum compatibility in the semiconductor industry. In the maglev system, the back electromotive force is inevitably generated when the system moves. The back electromotive force causes force/moment disturbances. Especially, the moment disturbances have negative effect on controlling the rotational motions (θ x , θ y , θ z ) whose control bandwidth is low. Therefore, the back electromotive force causes rotational motion errors. The rotational motion errors should be suppressed since they prevent high speed motion of the maglev system due to the rotational motion allowance of sensors. The rotational motion errors are suppressed by compensating the back electromotive force. In this paper, the back electromotive force, the cause of the rotational motion errors, is mathematically found in terms of the mover velocity and element of force-current matrix. A maglev system without the compensation was simulated and the rotational motion errors due to back electromotive force were found. Then, a feedforward compensated system using a mathematically modeled back electromotive force was simulated. It was verified that the feedforward compensation method with the modeled equation could be useful for suppressing rotational motion errors.

We integrated two-photon microscopy (TPM), second harmonic imaging microscopy (SHIM), near-infrared fluorescence microscopy (NIRF), confocal laser scanning microscopy (CLSM), and fluorescence lifetime imaging microscopy (FLIM) in one microscope system. Using our microscope system, five different images were obtained simultaneously.

Coronary arteries are covered by a thin layer of endothelial cells (ECs). Impairment of ECs is at the origin of coronary atherosclerosis and its clinical manifestations. However, the study of ECs in humans remains elusive because of a lack of an imaging tool with sufficient resolution. We have developed a light-based 1-µm-resolution microscopic imaging technology termed micro-optical coherence tomography (µOCT) that can be implemented in a coronary catheter. In this study, we investigated the capability of µOCT to visualize EC morphology. We stripped the endothelium from 36 fresh swine coronary segments with cyanoacrylate glue. Histology showed that the stripping procedure successfully removed ECs from the swine coronary arteries. Coronary segments were then imaged in 3D with µOCT, and were processed for histology and scanning electron microscopy (SEM). µOCT images of stripped vs. intact sites were volume rendered in 3D and visually compared. 3D-µOCT allowed visualization of EC pavementing on intact artery surfaces that was strongly correlated to that seen by SEM. EC pavementing disappeared, and surface roughness calculated by computed root mean squared error diminished significantly at the sites with stripped EC compared with intact sites. µOCT was also utilized in human cadaver coronary arteries, showing its capability of identifying EC morphology of human coronary plaque harboring leukocyte adhesion, EC stent strut tissue coverage, and lack of ECs at lesions with necrotic core or superficial nodular calcifications. In conclusion, µOCT enables EC visualization in coronary arteries, suggesting that it could be useful in patients with coronary artery disease to better understand the role of ECs in the pathogenesis of coronary artery disease.

Fluorescence lifetime imaging microscopy (FLIM), especially based on the most accurate and frequently used technique called time-correlated single photon counting (TCSPC), has been widely employed for it gives not only morphological information but also chemical information. Chemical properties such as pH value (Hanson et al., 2002), ion or oxygen concentration (Gilbert et al., 2007), molecular dynamics and even the disease progression (Park et al., 2012) can be investigated and studied by TCSPC-FLIM. This powerful traits of FLIM has led many researchers and developers to include FLIM into their imaging system. Even though TCSPC-FLIM technique gives precise and accurate measurement of fl uorescence decay curves, its core technology, called single photon counting (SPC), inherently limits the system from high speed imaging. In the case of the state-of-art single channel TCSPC-FLIM, more than one minute is required to produce an image of 512 by 512 pixels with 10% accuracy. The acquisition time could be lengthened up to five minutes for 3% accuracy (Kollner & Wolfrum, 1992). To overcome such drawback of the long acquisition time of TCSPC-FLIM, analog mean-delay FLIM (AMD-FLIM) was recently developed (Moon et al., 2009; Won et al., 2009). Rather than using the stochastic reconstruction used in SPC techniques, AMD method extracts a decay constant directly by subtracting the mean-delay time (or mean-arrival time) of reflected photon flux from that of fluorescence photon fl ux. The total amount of alterations in the mean-delay time

Aims Inflammation plays essential role in development of plaque disruption and coronary stent-associated complications. This study aimed to examine whether intracoronary dual-modal optical coherence tomography (OCT)-near-infrared fluorescence (NIRF) structural–molecular imaging with indocyanine green (ICG) can estimate inflammation in swine coronary artery. Methods and results After administration of clinically approved NIRF-enhancing ICG (2.0 mg/kg) or saline, rapid coronary imaging (20 mm/s pullback speed) using a fully integrated OCT-NIRF catheter was safely performed in 12 atheromatous Yucatan minipigs and in 7 drug-eluting stent (DES)-implanted Yorkshire pigs. Stronger NIRF activity was identified in OCT-proven high-risk plaque compared to normal or saline-injected controls ( P = 0.0016), which was validated on ex vivo fluorescence reflectance imaging. In vivo plaque target-to-background ratio (pTBR) was much higher in inflamed lipid-rich plaque compared to fibrous plaque ( P < 0.0001). In vivo and ex vivo peak pTBRs correlated significantly ( P < 0.0022). In vitro cellular ICG uptake and histological validations corroborated the OCT-NIRF findings in vivo . Indocyanine green colocalization with macrophages and lipids of human plaques was confirmed with autopsy atheroma specimens. Two weeks after DES deployment, OCT-NIRF imaging detected strong NIRF signals along stent struts, which was significantly higher than baseline ( P = 0.0156). Histologically, NIRF signals in peri-strut tissue co-localized well with macrophages. Conclusion The OCT-NIRF imaging with a clinical dose of ICG was feasible to accurately assess plaque inflammation and DES-related inflammation in a beating coronary artery. This highly translatable dual-modal molecular–structural imaging strategy could be relevant for clinical intracoronary estimation of high-risk plaques and DES biology.

Background and objective The tethered spectrally-encoded confocal endomicroscopy (SECM) capsule is an imaging device that once swallowed by an unsedated patient can visualize cellular morphologic changes associated with gastrointestinal (GI) tract diseases in vivo. Recently, we demonstrated a tethered SECM capsule for counting esophageal eosinophils in patients with eosinophilic esophagitis (EoE) in vivo. Yet, the current tethered SECM capsule is far too long to be widely utilized for imaging pediatric patients, who constitute a major portion of the EoE patient population. In this paper, we present a new tethered SECM capsule that is 33% shorter, has an easier and repeatable fabrication process, and produces images with reduced speckle noise. Materials and methods The smaller SECM capsule utilized a miniature condenser to increase the fiber numerical aperture and reduce the capsule length. A custom 3D-printed holder was developed to enable easy and repeatable device fabrication. A dual-clad fiber (DCF) was used to reduce speckle noise. Results The fabricated SECM capsule (length = 20 mm; diameter = 7 mm) had a similar size and shape to a pediatric dietary supplement pill. The new capsule achieved optical sectioning thickness of 13.2 μm with a small performance variation between devices of 1.7 μm. Confocal images of human esophagus obtained in vivo showed the capability of this new device to clearly resolve microstructural epithelial details with reduced speckle noise. Conclusions We expect that the smaller size and better image performance of this new SECM capsule will greatly facilitate the clinical adoption of this technology in pediatric patients and will enable more accurate assessment of EoE-suspected tissues. Lasers Surg. Med. 9999:XX-XX, 2018. © 2019 Wiley Periodicals, Inc.

Magnetic levitation planar motor was researched and developed for vacuum compatibility. Laser interferometers are used in order for high precision measurement. However, the magnetic levitations cannot experience full working range due to the high sensitivity of the laser interferometers. Especially, angular alignment is important for laser interferometer to measure the motions. In this paper, a cause of the lower angular alignment along the long range motion is investigated and feedforward reference compensation using bilinear interpolation is utilized to improve the angular alignment. In order for verifying the compensation, experimental work was performed and the result showed that the angular alignment was improved. Finally, the magnetic levitation planar motor can have full working range motions including 2-dimensional motions by suggested compensation.

High internal phase emulsions have been widely used as templates for various porous materials, but special strategies are required to form, in particular, particle-covered ones that have been more difficult to obtain. Here, we report a versatile strategy to produce a stable high internal phase Pickering emulsion by exploiting a depletion interaction between an emulsion droplet and a particle using water-soluble polymers as a depletant. This attractive interaction facilitating the adsorption of particles onto the droplet interface and simultaneously suppressing desorption once adsorbed. This technique can be universally applied to nearly any kind of particle to stabilize an interface with the help of various non- or weakly adsorbing polymers as a depletant, which can be solidified to provide porous materials for many applications., The fabrication of emulsion droplets stabilized by solid particles adsorbed on the interface is restricted to delicate interfacial conditions. Here, Kim et al. show a general approach to prepare them using the depletion interaction, modified by soluble polymers, between particles and emulsions.

Impairment of endothelial cells (ECs) is at the origin of coronary atherosclerosis. More recently, plaque erosion has emerged as the second most prevalent pathological finding in acute coronary syndrome (ACS). However, the study of ECs in humans remains elusive because of a lack of an imaging tool

Introduction: Atherosclerosis is a process of lipid accumulation and inflammation. In particular, plaque macrophages are associated with fibrous cap destabilization and rupture. We hypothesized that the specific delivery of a PPAR-γ agonist to inflamed plaques via nanoprobe (NP) targeting macrophage mannose receptors could reduce plaque burden and inflammation. Methods and Results: Macrophage mannose receptor targetable nanoprobe (MMR-NP) was fabricated by chemically conjugating thiolated glycol chitosan with mannose-PEG-maleimide, followed by incorporating PPAR-γ agonist (lobeglitazone) into this nanoscale delivery system (MMR-Lobe). For in vivo monitoring of therapeutic response, near-infrared fluorescence NP was prepared by conjugation of Cy5.5 (ext/emi 675/694 nm) with MMR-NP. In cellular uptake study, the MMR-NP showed high affinity to mannose receptors on macrophages. MMR-Lobe attenuated LPS-induced inflammatory cytokines such as TNF-α, IL-6, and MMP-9 in RAW264.7 cells. Additionally, MMR-Lobe increased expression of ABCA1, ABCG1 and LXR-α, known as PPAR-γ regulator genes involved in cholesterol efflux, in RAW264.7 cells. Using a customized high-resolution intravital fluorescence microscope, the in vivo serial imaging of carotid atheroma in apoE-/- mice injected with MMR-Lobe (7 mg/kg, twice weekly for 4 weeks) revealed a significant decrease in plaque burden and inflammation (Figure) as compared to baseline, or non-treated controls, or p.o. Lobe treated mice (p Conclusions: MMR-Lobe was able to selectively target atheroma macrophages, and effectively reduce both plaque burden and inflammation as assessed by serial intravital optical imaging. We suggest that nanoprobe-mediated PPAR-γ agonist delivery targeting plaque macrophages holds a promising theranostic approach for high-risk atheromata.

Background: Reactive oxygen species (ROS) play a central role in cigarette smoking-induced atherogenesis. The present study aims to assess the smoking-induced acute oxidative stress within vasculatures, and evaluates whether the resveratrol, a natural polyphenol antioxidant, can counteract this ROS production, using a customized, high resolution intravital optical imaging in real-time. Methods and Results: 20-week-old male C57BL/6 mice were divided into four groups according to the preceding administration of resveratrol (R) (25mg/kg via gavage, for 7 days) and exposure to cigarette smoke (CS). To in vivo assess acute oxidative stress in blood vessels, dihydroethidium, which forms a red fluorescence (ethidium, excitation/emission: 520nm/610nm) upon reaction with ROS, was injected intraperitoneally. During CS exposure, temporal changes of fluorescence signals from the mouse cremaster muscle including vasculatures were assessed by intravital optical imaging for 15 minutes. Fluorescence signals were much more pronounced in CS exposed mice than controls (p Conclusions: The intravital optical imaging was able to in vivo estimate the dynamic changes of ROS production by CS exposure. Our data demonstrated that even a brief exposure to CS increased oxidative stress in vasculatures promptly, and the resveratrol exerts protective effects against the CS-induced acute oxidative stress.

Macrophages mediate atheroma expansion and disruption, and denote high-risk arterial plaques. Therefore, they are substantially gaining importance as a diagnostic imaging target for the detection of rupture-prone plaques. Here, we developed an injectable near-infrared fluorescence (NIRF) probe by chemically conjugating thiolated glycol chitosan with cholesteryl chloroformate, NIRF dye (cyanine 5.5 or 7), and maleimide-polyethylene glycol-mannose as mannose receptor binding ligands to specifically target a subset of macrophages abundant in high-risk plaques. This probe showed high affinity to mannose receptors, low toxicity, and allowed the direct visualization of plaque macrophages in murine carotid atheroma. After the scale-up of the MMR-NIRF probe, the administration of the probe facilitated in vivo intravascular imaging of plaque inflammation in coronary-sized vessels of atheromatous rabbits using a custom-built dual-modal optical coherence tomography (OCT)-NIRF catheter-based imaging system. This novel imaging approach represents a potential imaging strategy enabling the identification of high-risk plaques in vivo and holds promise for future clinical implications.

A fluorescence lifetime imaging microscopy (FLIM) integrated with two-photon excitation technique was developed. A wavelength-tunable femtosecond pulsed laser with nominal pulse repetition rate of 76-MHz was used to acquire FLIM images with a high pixel rate of 3.91 MHz by processing the pulsed two-photon fluorescence signal. Analog mean-delay (AMD) method was adopted to accelerate the lifetime measurement process and to visualize lifetime map in real-time. As a result, rapid tomographic visualization of both structural and chemical properties of the tissues was possible with longer depth penetration and lower photo-damage compared to the conventional single-photon FLIM techniques.

Introduction: Plaque inflammation is associated with fibrous cap destabilization and rupture. Hypothesis: We hypothesized that the specific delivery of a novel PPARγ agonist, lobeglitazone, to inflamed plaques via nanoparticle (NP) targeting mannose receptors on macrophages could reduce inflammation in atheromata and thus, tested it using serial optical intravital molecular imaging. Methods and Results: A novel near-infrared fluorescence (NIRF) probe was fabricated by chemically conjugating thiolated glycol chitosan with mannose-PEG, and NIRF dye Cy5.5 (ext/emi 675/694 nm) for imaging. Lobeglitazone was incorporated into NIRF-NP targeting macrophage mannose receptors (MMR). In vitro, MMR-NIRF-NP was selectively uptaken by RAW264.7 cells, and NIRF signals were estimated to be 10-fold over control group (p Conclusions: Lobeglitazone-incorporated MMR-NIRF-NP was able to selectively target and image atheroma macrophages, and effectively reduce plaque inflammation as assessed by intravital optical imaging. NP-mediated lobeglitazone delivery could be a promising therapeutic strategy for high-risk plaque stabilization.

FLIM (Fluorescence Lifetime Imaging Microscopy) has been considered an effective technique to investigate chemical properties of the specimens, especially of biological samples. Despite of this advantageous trait, researchers in this field have had difficulties applying FLIM to their systems because acquiring an image using FLIM consumes too much time. To increase the FLIM speed, many methodologies have been developed and applied to the system. One of the recent methodologies is an analogue mean delay based FLIM using a PMT and digitizer for image reconstruction. In this system, however, imaging time is largely dependent upon several parameters such as data transfer rate, sampling rate of an A/D converter, and signal width etc. In this paper, such parametric optimization method is introduced for faster acquisition of the image.

Fluorescence lifetime imaging microscopy (FLIM) is a powerful technique to visualize photophysical characteristics of biological targets. However, conventional FLIM methods have some limitations that restrict obtaining high-precision images in real time. Here, we propose a high-speed time-resolved laser-scanning microscopy by incorporating a novel line-to-pixel referencing method into the previously suggested analog mean-delay (AMD) method. The AMD method dramatically enhances the photon accumulation speed for achieving the certain precision compared to the time-correlated single-photon counting (TCSPC) method while maintaining high photon efficiency. However, its imaging pixel rate can still be restricted by the rearm time of the digitizer when it is triggered by laser pulses. With our line-to-pixel referencing method, the pulse train repeats faster than the trigger rearm time can be utilized by generating a line trigger, which is phase-locked with only the first pulse in each horizontal line composing an image. Our proposed method has been tested with a pulsed laser with 40 MHz repetition rate and a commercial digitizer with a 500 ns trigger rearm time, and a frame rate of 3.73 fps with a pixel rate of 3.91 MHz was accomplished while maintaining the measurement precision under 20 ps.

During the research using fluorescence-tagged or auto-fluorescence molecules, meaningful information is often buried deep inside the tissue, not its surface. Therefore, especially in the field of biomedical imaging, acquiring optically sectioned images from deep inside the tissue is very important. As well know already, confocal laser scanning microscopy (the most well-known optical sectioning microscopy) gives axially-resolved fluorescence information using the physical background blocking component called pinhole. However, the axial range of imaging is practically limited due to such optical phenomena as the light scattered and absorbed in the tissue. However, nonlinear optical microscopy (e.g. Multiphoton microscopy, harmonic generation microscopy, coherent anti-Stokes Raman spectroscopy) realized by the development of ultrafast light sources has been used for visualizing various tissues, especially in vivo, because of their low sensitivity to the limitation caused by the scattering and the absorption of light. Although nonlinear optical microscopy gives deep tissue image, it is not easy for many researcher to build customized nonlinear system. Here, we introduce an easy and simple way designing and developing such nonlinear optical microscope with upright or inverted epi-illumination platform using commercial optical components only.