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Alkaline phosphatase (ALP) is a membrane-bound enzyme widely present in biological tissues and is extensively used as a biomarker in clinical diagnosis because abnormal ALP levels in organisms are often closely related to many diseases. In order to meet the urgent needs of human health and the development of precision medicine, the classic method of detecting ALP seems to be unable to meet a "tailor-made" treatment plan for patients. In order to get the best therapeutic effect and the lowest side effects, the researchers have made considerable efforts and established numerous strategies for detecting ALP. In this review, we focus on the current development and basic principles of fluorescence strategy to detect ALP, and compare the advantages and disadvantages of detecting ALP based on different fluorescent detection modes. In addition, to more intuitively understand the performance of different materials for detecting ALP, a table is presented. Finally, the future trends and prospective in this field will be discussed, it could be speculated that invent and design the detection mechanism based on the relationship between the new material and ALP is the key to effective detection of ALP, and process data with a computer analog digital platform will be a welcome and obsessive technology. It is hoped that some insights and inspiration for future research work will be provided., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: To compare the keratometric measurements of the Verion image guided system with the partial coherence interferometry (IOLMaster700) and the scheimpflug corneal topography (Pentacam). Methods: In this cross-sectional study, 146 eyes of 88 patients [mean age, (69±10) years; 62 eyes of male patients and 84 eyes of female patients] with cataract were examined before cataract extraction and IOL implantation surgery in the Tenth People's Hospital Affiliated to Tongji University from January 2016 to March 2017. Corneal curvature values acquired by the Verion optical imaging system were compared with the IOLMaster700 and the Pentacam. Keratometric data, magnitude of astigmatism, and steep astigmatic axis measurements from all three instruments were also compared. According to the steep axis of astigmatism, the eyes were divided into groups of with the rule (WTR), against the rule (ATR), and oblique astigmatism. The parameter differences between the Verion and the other devices in the three groups were analyzed. The data were evaluated using repeated measures analysis of variance and Bland-Altman plots. Results: The values of steep keratometry (Ks) of the Verion, IOLMaster700, and Pentacam were (44.60±1.32), (44.73±1.37), (44.43±1.35) D, respectively. The values of flat keratometry (Kf) of the Verion, IOLMaster700, and Pentacam were (43.51±1.41), (43.51±1.52), (43.40±1.45) D, respectively. The values of mean keratometry (Km) of the Verion, IOLMaster700, and Pentacam were (44.60±1.32), (44.12±1.35), (43.92±1.34) D, respectively. The values of astigmatism magnitude were (1.09±0.87), (1.24±1.02), (1.04±0.80) D, respectively. The F values were 1.81, 0.31, 0.93 and 2.22 in relation to Ks, Kf, Km and corneal astigmatism magnitude among the three instruments, respectively (all P >0.05). The 95% confidence intervals of Kf, Ks and astigmatism magnitude between the Verion and the other two devices were -1.10 to 0.80 D, -0.55 to 0.88 D; -1.10 D to 1.10 D, -0.80 to 1.04 D; -1.21 to 0.90 D, -0.92 to 1.02 D. The outcomes were considered acceptable. There were no statistically differences in steep astigmatic axis measurements between the Verion and the other two instruments among the WTR (62 eyes), ATR (45 eyes) and oblique (15 eyes) groups ( F= 0.63, 0.37; P= 0.54, 0.72, respectively). Conclusions: The Verion image guided system is a reliable system for the measurement of keratometry values and astigmatism. The keratometric power, magnitude and steep axis of astigmatism have no significant difference, and there is a good agreement among the Verion, IOL Master 700, and Pentacam. (Chin J Ophthalmol, 2020, 56: 47 - 52) .

The alarming dynamics of antibiotic-resistant infections calls for the development of rapid and point-of-care (POC) antibiotic susceptibility testing (AST) methods. Here, we demonstrated the first completely stand-alone microfluidic system that allowed the execution of digital enumeration of bacteria and digital antibiograms without any specialized microfluidic instrumentation. A four-chamber gravity-driven step emulsification device generated ∼2000 monodisperse 2 nanoliter droplets with a coefficient of variation of 8.9% of volumes for 95% of droplets within less than 10 minutes. The manual workload required for droplet generation was limited to the sample preparation, the deposition into the sample inlet of the chip and subsequent orientation of the chip vertically without an additional pumping system. The use of shallow chambers imposing a 2D droplet arrangement provided superior stability of the droplets against coalescence and minimized the leakage of the reporter viability dye between adjacent droplets during long-term culture. By using resazurin as an indicator of the growth of bacteria, we were also able to reduce the assay time to ∼5 hours compared to 20 hours using the standard culture-based test.

Objective: Fluorescence molecular imaging (FMI) has emerged as a promising tool for surgical guidance in oncology, with one of the few remaining challenges being the ability to offer quality control and data referencing. This paper investigates the use of a novel composite phantom to correct and benchmark FMI systems., Methods: This paper extends on previous work by describing a phantom design that can provide a more complete assessment of FMI systems through quantification of dynamic range and determination of spatial illumination patterns for both reflectance and fluorescence imaging. Various performance metrics are combined into a robust and descriptive "system benchmarking score," enabling not only the comprehensive comparison of different systems, but also for the first time, correction of the acquired data., Results: We show that systems developed for targeted fluorescence imaging can achieve benchmarking scores of up to 70%, while clinically available systems optimized for indocyanine green are limited to 50%, mostly due to greater leakage of ambient and excitation illumination and lower resolution. The image uniformity can also be approximated and employed for image flat-fielding, an important milestone toward data referencing. In addition, we demonstrate composite phantom use in assessing the performance of a surgical microscope and of a raster-scan imaging system., Conclusion: Our results suggest that the new phantom has the potential to support high-fidelity FMI through benchmarking and image correction., Significance: Standardization of the FMI is a necessary process for establishing good imaging practices in clinical environments and for enabling high-fidelity imaging across patients and multi-center imaging studies.

Background/aims: To compare white light imaging (WLI) with linked color imaging (LCI) and blue LASER imaging (BLI) in endoscopic findings of Helicobacter pylori presently infected, previously infected, and uninfected gastric mucosae for visibility and inter-rater reliability., Methods: WLI, LCI and BLI bright mode (BLI-bright) were used to obtain 1,092 endoscopic images from 261 patients according to the Kyoto Classification of Gastritis. Images were evaluated retrospectively by 10 experts and 10 trainee endoscopists and included diffuse redness, spotty redness, map-like redness, patchy redness, red streaks, intestinal metaplasia, and an atrophic border (52 cases for each finding, respectively). Physicians assessed visibility as follows: 5 (improved), 4 (somewhat improved), 3 (equivalent), 2 (somewhat decreased), and 1 (decreased). Visibility was assessed from totaled scores. The inter-rater reliability (intraclass correlation coefficient) was also evaluated., Results: Compared with WLI, all endoscopists reported improved visibility with LCI: 55.8% for diffuse redness; LCI: 38.5% for spotty redness; LCI: 57.7% for map-like redness; LCI: 40.4% for patchy redness; LCI: 53.8% for red streaks; LCI: 42.3% and BLI-bright: 80.8% for intestinal metaplasia; LCI: 46.2% for an atrophic border. For all endoscopists, the inter-rater reliabilities of LCI compared to WLI were 0.73-0.87., Conclusion: The visibility of each endoscopic finding was improved by LCI while that of intestinal metaplasia was improved by BLI-bright., (© 2019 S. Karger AG, Basel.)

Microcirculation plays a crucial role in delivering oxygen and nutrients to living tissues and in removing metabolic wastes from the human body. Monitoring the velocity of blood flow in microcirculation is essential for assessing various diseases, such as diabetes, cancer, and critical illnesses. Because of the complex morphological pattern of the capillaries, both In-vivo capillary identification and blood flow velocity measurement by conventional optical capillaroscopy are challenging. Thus, we focused on developing an In-vivo optical microscope for capillary imaging, and we propose an In-vivo full-field flow velocity measurement method based on intelligent object identification. The proposed method realizes full-field blood flow velocity measurements in microcirculation by employing a deep neural network to automatically identify and distinguish capillaries from images. In addition, a spatiotemporal diagram analysis is used for flow velocity calculation. In-vivo experiments were conducted, and the images and videos of capillaries were collected for analysis. We demonstrated that the proposed method is highly accurate in performing full-field blood flow velocity measurements in microcirculation. Further, because this method is simple and inexpensive, it can be effectively employed in clinics.

In this paper, we present a novel hyperspectral system designed for imaging the human iris in vivo. The instrument is based on a tunable light source (from 480 nm to 900 nm) and a monochrome camera. The system can acquire hyperspectral cubes in 4 s, with spatial and spectral resolutions of 11 μm and 20 nm, respectively. The optical characterization of the new instrument is described, and we demonstrate its safety with respect to ANSI standards. Preliminary in vivo measurements in human volunteers are also presented.

We report an application for the smartphone as an accurate and unbiased reading platform of a lateral flow immunoassays for food safety application. In particular, this report focuses on detection of food-borne bacteria in samples extracted from food matrices such as ground beef and spinach. The lateral flow assay is a widely accepted methodology owing to its on-site results, low-cost analysis, and ease of use with minimum user inputs, even though sensitivity is not quite equivalent to that of standard laboratory equipment. An antibody-antigen relationship is transduced into a color change on a nitrocellulose pad while visual interpretation of this color change can result in uncertainty, particularly near the detection limit of the assay. Employing the high resolution integrated camera, constant illumination from light source, and computing power of a smartphone, we provide an objective and accurate method to determine the bacterial cell concentration in a food matrix based on the regression model from the color intensity of test lines. A 3D-printed sample holder was designed for representative commercial lateral flow assays and an in-house application was developed in Android Studio to solve the inverse problem to provide cell concentration information from the color intensity. Test results with E.coli O157:H7 as a model organism suggests that smartphone-based reader can detect 10 4 -10 5 CFU/ml from ground beef and spinach food matrices., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Plant embryogenesis begins with fertilization and ends with the generation of the basic body plan of the future plant. Despite its importance, the dynamics of flowering plant ontogeny have long been a mystery, because the embryo develops deep in the maternal tissue. Recently, an embryonic live-cell imaging system was established in Arabidopsis thaliana by developing an in vitro ovule cultivation method and utilizing two-photon excitation microscopy (2PEM), which is suitable for deep imaging. This system enabled us to visualize intracellular dynamics during zygote polarization and monitor the cell division pattern during embryogenesis from the zygote until organ formation. In this chapter, we describe a method that allows for high-resolution imaging of cytoskeletal rearrangements in the zygote and long-term tracing of embryo patterning.

Significance : Spatial frequency domain imaging (SFDI) is a diffuse optical measurement technique that can quantify tissue optical absorption (μ a ) and reduced scattering ( μ s ' ) on a pixel-by-pixel basis. Measurements of μ a at different wavelengths enable the extraction of molar concentrations of tissue chromophores over a wide field, providing a noncontact and label-free means to assess tissue viability, oxygenation, microarchitecture, and molecular content. We present here openSFDI: an open-source guide for building a low-cost, small-footprint, three-wavelength SFDI system capable of quantifying μ a and μ s ' as well as oxyhemoglobin and deoxyhemoglobin concentrations in biological tissue. The companion website provides a complete parts list along with detailed instructions for assembling the openSFDI system.

Aim : We describe the design of openSFDI and report on the accuracy and precision of optical property extractions for three different systems fabricated according to the instructions on the openSFDI website.

Approach : Accuracy was assessed by measuring nine tissue-simulating optical phantoms with a physiologically relevant range of μ a and μ s ' with the openSFDI systems and a commercial SFDI device. Precision was assessed by repeatedly measuring the same phantom over 1 h.

Results : The openSFDI systems had an error of 0  ±  6  %   in μ a and -2  ±  3  %   in μ s ' , compared to a commercial SFDI system. Bland-Altman analysis revealed the limits of agreement between the two systems to be   ±  0.004  mm   -  1 for μ a and -0.06 to 0.1  mm   -  1 for μ s ' . The openSFDI system had low drift with an average standard deviation of 0.0007  mm   -  1 and 0.05  mm   -  1 in μ a and μ s ' , respectively.

Conclusion : The openSFDI provides a customizable hardware platform for research groups seeking to utilize SFDI for quantitative diffuse optical imaging.

Bioimaging instrumentation can be used to observe environmental phenomena such as the transport, retention, and distribution of bacteria in soils in situ in a real-time, nondestructive manner. Bacteria designed to emit bioluminescence light signals are injected into a transparent column packed with soils, and then the column is placed into a bioimaging instrument, such as a PerkinElmer IVIS Spectrum, while it is connected through thin teflon tubes to other parts of the column system located outside of the imaging chamber, including a fraction collector for collecting effluent solution and a pump for introducing bacterial suspension or experimental solution. After self-correction of soil autofluorescence and bioluminescence and setup of required imaging parameters, the transport experiment is initiated by introducing the bacterial suspension to the soil column while the spatiotemporal distribution of bioluminescent bacteria in the entire soil column is imaged. Finally, the images are processed to analyze bacterial migration in the soil under various environmental conditions in comparison with the breakthrough and elution curves of the bacteria obtained by analyzing the effluent samples.

In this paper, we have investigated multi-channel switching of light incidence in multiple directions to improve image clarity in surface plasmon microscopy (SPM) for robust and consistent imaging performance regardless of the pattern geometry and shape. Multi-channel light switching in SPM allows significant reduction of adverse scattering effects by surface plasmon (SP). For proof of concept, an eight-channel spatially switched SPM (ssSPM) system has been set up. The results with reference objects including square arrays and Siemens stars experimentally confirm much improved images with ssSPM by reducing the artifacts of SP scattering significantly. On a quantitative basis, contrast analysis preformed with square arrays shows image contrast enhanced by more than three times over conventional SPM. Three image reconstruction algorithms were evaluated for optimal image acquisition. It is suggested that averaging combined with minimum-filtering produces the highest resolution. ssSPM was applied to label-free imaging of primary neuron cultures and shown to present enhanced images with clarity far better than conventional SPM., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Objective: Diagnostics which provide objective information to facilitate evidence-based treatment decisions could improve the chance of wound healing. Accurate wound measurements, objective bacterial assessment, and the regular, consistent tracking of these parameters are important aspects of wound care. This study aimed to assess the accuracy, clinical incorporation and documentation capabilities of a handheld bacterial fluorescence imaging device (MolecuLight i :X)., Method: Benchtop wound models with known dimensions and clinical wound images were repeatedly measured by trained clinicians to quantify accuracy and intra/inter-user coefficients of variation (COV) of the imaging device measurement software. In a clinical trial of 50 wounds, wound dimensions were digitally measured and fluorescence images were acquired to assess for the presence of bacteria at moderate-to-heavy loads. Finally, fluorescence imaging was implemented into the routine assessment of 22 routine diabetic foot ulcers (DFU) to determine appropriate debridement level and location based on bacterial fluorescence signals., Results: Wound measurement accuracy was >95% (COV <3%). In the clinical trial of 50 wounds, 72% of study wounds demonstrated positive bacterial fluorescence signals. Levine sampling of wounds was found to under-report bacterial loads relative to fluorescence-guided curettage samples. Furthermore, fluorescence documentation of bacterial presence and location(s) resulted in more aggressive, fluorescence-targeted debridement in 17/20 DFUs after standard of care debridement failed to eliminate bacterial fluorescence in 100% of DFU debridements., Conclusion: The bacterial fluorescence imaging device can be readily implemented for objective, evidenced-based wound assessment and documentation at the bedside. Bedside localisation of regions with moderate-to-heavy bacterial loads facilitated improved sampling, debridement targeting and improved wound bed preparation.

Phase and polarization of coherent light are highly perturbed by interaction with microstructural changes in premalignant tissue, holding promise for label-free detection of early tumors in endoscopically accessible tissues such as the gastrointestinal tract. Flexible optical multicore fiber (MCF) bundles used in conventional diagnostic endoscopy and endomicroscopy scramble phase and polarization, restricting clinicians instead to low-contrast amplitude-only imaging. We apply a transmission matrix characterization approach to produce full-field en-face images of amplitude, quantitative phase, and resolved polarimetric properties through an MCF. We first demonstrate imaging and quantification of biologically relevant amounts of optical scattering and birefringence in tissue-mimicking phantoms. We present an entropy metric that enables imaging of phase heterogeneity, indicative of disordered tissue microstructure associated with early tumors. Finally, we demonstrate that the spatial distribution of phase and polarization information enables label-free visualization of early tumors in esophageal mouse tissues, which are not identifiable using conventional amplitude-only information.

Significance : While clinical treatment of actinic keratosis by photodynamic therapy (PDT) is widely practiced, there is a well-known variability in response, primarily caused by heterogeneous accumulation of the photosensitizer protoporphyrin IX (PpIX) between patients and between lesions, but measurement of this is rarely done. Aim : Develop a smartphone-based fluorescence imager for simple quantitative photography of the lesions and their PpIX levels that can be used in a new clinical workflow to guide the reliability of aminolevulinic acid (ALA) application for improved lesion clearance. Approach : The smartphone fluorescence imager uses an iPhone and a custom iOS application for image acquisition, a 3D-printed base for measurement standardization, an emission filter for PpIX fluorescence isolation, and a 405-nm LED ring for optical excitation. System performance was tested to ensure measurement reproducibility and the ability to capture photosensitizer accumulation and photobleaching in pre-clinical and clinical settings. Results : PpIX fluorescence signal from tissue-simulating phantoms showed linear sensitivity in the 0.01 to 2.0 μM range. Murine studies with Ameluz® aminolevulinic acid (ALA) gel and initial human testing with Levulan® ALA cream verified that in-vivo imaging was feasible, including that PpIX production over 1 h is easily captured and that photobleaching from the light treatment could be quantified. Conclusions : The presented device is the first quantitative wide-field fluorescence imaging system for PDT dosimetry designed for clinical skin use and for maximal ease of translation into clinical workflow. The results lay the foundation for using the system in clinical studies to establish treatment thresholds for the individualization of PDT treatment.

Intravenous indocyanine green (ICG) has been reported to localize intra-abdominal metastatic lesions in several clinical trials. Our pilot study aimed to investigate the feasibility and safety of ICG fluorescence localization in pulmonary metastasectomy using a near-infrared fluorescence thoracoscope. Each patient received intravenous 0.25 or 0.5 mg/kg of ICG. The maximum diameter of the tumor on computed tomography ranged from 0.5 to 3.5 (median: 1.15) cm. Intravenous ICG injection localized pulmonary metastases in a portion (3 patients) of the enrolled patients. Our preliminary results provided us with important information to modify the study protocol., Competing Interests: None., (Georg Thieme Verlag KG Stuttgart · New York.)

Objective: Currently, neurosurgical vascular surgery frequently uses indocyanine green (ICG)-videography (VG) to evaluate the blood flow in brain vessels. Although ICG-VG delineates intravascular ICG fluorescence as a high-intensity signal in gray-scale with dark background, it is hard to identify anatomical structures, including vasculature or surgical devices simultaneously. This report developed combination of a near-infrared (NIR) camera with particular sensitivity and an optical filter to observe the blood-flow conditions and anatomical structures., Methods: To overcome the specific issues of ICG-VG, we applied a high-sensitivity camera with a 980-nm NIR component to delineate anatomical and fluorescence images, detecting signals between 830 and 1000 nm simultaneously during operation. We used a diluted ICG phantom to evaluate fluorescence signal changes by changing wavelength of the emission light. For clinical use, we used a high-sensitivity NIR camera with a high-pass filter on a surgical microscope. The new NIR system detected signals between 770 and 1000 nm, and the lighting system illuminated objects mainly at 980-nm wavelength. Both images with the blood flow and anatomical structures were projected to the smart glasses in real time., Results: In the phantom experiment, we found that the emission light with wide band widths (575-800 nm) evoked various intensities of ICG fluorescence. This new NIR system allowed us to observe ICG fluorescence and anatomical structures without image fusion or time-delay. The both information of anatomy and fluorescence was projected on wearable smart glasses. Furthermore, the new NIR system detected ICG-fluorescence signals for a longer duration than the original camera, which allowed us to achieve careful and detailed observation of more vasculature and fine vessels., Conclusions: This study proposes a new NIR system and emphasizes simultaneous observation of anatomy and fluorescence signals during operation. It paves the way for further possibilities in the development of optical systems. To understand the natural phenomena and combination of different scientific and clinical fields, it might be important to understand and combine not only fluorescence, but also natural science, optics, and background pathology. This simple system would be available for neuroendoscope and robotic surgery., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Purpose: Urinary stone treatment has been strongly influenced by advances in technology. Nevertheless, the photonic characteristics of stones as the treatment target have been neglected. Monitoring fluorescence spectra is sufficient for automatic target differentiation and laser feedback control as previously described. We investigated the characteristics of fluorescence signals and the clinical practicability of real-time laser feedback control during lithotripsy., Materials and Methods: Fluorescence excitation light was superimposed on a holmium laser beam into the treatment fiber. Spectra were recorded and signal amplitude changes were analyzed during increases in distance between the fiber tip and the stone to identify the optimal threshold level for stone recognition. Ho:YAG lithotripsy was performed under in vitro surgical conditions in porcine tissue while our feedback system autonomously controlled the laser impulse release during lithotripsy. The tissue was then endoscopically and macroscopically examined for laser induced lesions., Results: Mean ± SD autofluorescence signal amplitudes from urinary stone samples varied between 142 ± 29 and 1,521 ± 152 ADU while tissue and endoscope coating emission was negligible. Signal amplitude decreased rapidly at distances larger than 1 to 2 mm. Clinically reliable threshold values for target recognition could be set to prevent laser pulse emission if the stone was out of range or urothelial tissue might be harmed by laser irradiation. We observed no incorrectly released laser pulse or injury to tissue during autonomously controlled holmium laser lithotripsy., Conclusions: Our laboratory study strengthens the evidence that tracking real-time autofluorescence spectra during endoscopic stone surgery via automatic feedback control of the laser impulse release may become a potentially useful clinical tool for surgeons who navigate in the upper urinary tract.

Fluorescence imaging, as a commonly used scientific tool, is widely applied in various biomedical and material structures through visualization technology. Highly selective and sensitive luminescent biological probes, as well as those with good water solubility, are urgently needed for biomedical research. In contrast to the traditional aggregation-caused quenching of fluorescence, in the unique phenomenon of aggregation-induced emission (AIE), the individual luminogens have extremely weak or no emissivity because they each have free intramolecular motion; however, when they form aggregates, these components immediately "light up". Since the discovery of "turn-on" mechanism, researchers have been studying and applying AIE in a variety of fields to develop more sensitive, selective, and efficient strategies for the AIE dyes. There are numerous advantages to the use of AIE-based methods, including low background interference, strong contrast, high performance in intracellular imaging, and the ability for long-term monitoring in vivo. In this review, two typical examples of AIEgens, TPE-Cy and TPE-Ph-In, are described, including their structure properties and applications. Recent progress in the biological applications is mainly focused on. Undoubtedly, in the near future, an increasing number of encouraging and practical ideas will promote the development of more AIEgens for broad use in biomedical applications., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background and Objective: Microcirculation plays a critical role in physiologic processes and several disease states. Laser speckle imaging (LSI) is a full-field, real-time imaging technique capable of mapping microvessel networks and providing relative flow velocity within the vessels. In this study, we demonstrate that LSI combine with multispectral reflectance imaging (MSRI), which allows for distinction between veins and arteries in the vascular flow maps produced by LSI. We apply this combined technique to mouse cerebral vascular network in vivo, comparing imaging through the skull, to the dura mater and brain directly through a craniectomy, and through a transparent cranial "Window to the Brain" (WttB) implant., Study Design/materials and Methods: The WttB implant used in this study is made of a nanocrystalline Yttria-Stabilized-Zirconia ceramic. MSRI was conducted using white-light illumination and filtering the reflected light for 560, 570, 580, 590, 600, and 610 nm. LSI was conducted using an 810 nm continuous wave near-infrared laser with incident power of 100 mW, and the reflected speckle pattern was captured by a complementary metal-oxide-semiconductor (CMOS) camera., Results: Seven vessel branches were analyzed and comparison was made between imaging through the skull, craniectomy, and WttB implant. Through the skull, MSRI did not detect any vessels, and LSI could not image microvessels. Imaging through the WttB implant, MSRI was able to identify veins versus arteries, and LSI was able to image microvessels with only slightly higher signal-to-noise ratio and lower sharpness than imaging the brain through a craniectomy., Conclusions: This study demonstrates the ability to perform MSRI-LSI across a transparent cranial implant, to allow for cerebral vascular networks to be mapped, including microvessels. These images contain additional information such as vein-artery separation and relative blood flow velocities, information which is of value scientifically and medically. The WttB implant provides substantial improvements over imaging through the murine cranial bone, where microvessels are not visible and MSRI cannot be performed. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc., (© 2019 Wiley Periodicals, Inc.)

Recently, many reconstruction methods have been developed to improve the lateral resolution of acoustic-resolution photoacoustic microscopy (ARPAM) in out-of-focus regions. Though these methods enhance image resolution to some extent, they require advanced computational hardware and large computational time, especially for three-dimensional (3-D) cases. However, some methods do not consider the finite size of a transducer, while others employ numerical discretization to build a focused transducer model that is less efficient and accurate. To overcome these problems, we propose a 3-D ARPAM imaging reconstruction method with high precision, high efficiency, and low memory cost. It inherits the framework of model-based reconstructions and incorporates the forward acoustic model in the hybrid domain. This hybrid-domain acoustic model promotes an analytical solution to establish a focused transducer model. Furthermore, the non-uniform fast Fourier transform (NUFFT) and deconvolution methods are introduced to reduce the required computational time and memory volume for 3-D reconstructions. According to the experimental results reconstructed by the proposed method, the lateral resolution of an ARPAM image recorded by a 20-MHz focused transducer (NA 0.393) can reach 88.39 [Formula: see text]. This resolution exceeds the diffraction limitation of the focused transducer ( [Formula: see text]). When reconstructing a 3-D image with 200×200×150 pixels, the proposed method takes only 8.15 s using a laptop loaded with Intel Core i7-8550U CPU at 1.8 GHz and 1.06-GB memory.

The excited state lifetime of a fluorophore together with its fluorescence emission spectrum provide information that can yield valuable insights into the nature of a fluorophore and its microenvironment. However, it is difficult to obtain both channels of information in a conventional scheme as detectors are typically configured either for spectral or lifetime detection. We present a fiber-based method to obtain spectral information from a multiphoton fluorescence lifetime imaging (FLIM) system. This is made possible using the time delay introduced in the fluorescence emission path by a dispersive optical fiber coupled to a detector operating in time-correlated single-photon counting mode. This add-on spectral implementation requires only a few simple modifications to any existing FLIM system and is considerably more cost-efficient compared to currently available spectral detectors.

Background: To utilize plaque scoring indices as objective measures, a certain standard of intra-rater and inter-rater agreement must be met. Achievement of high consistency among several examiners in the assessment of images from Q-ray cam (for quantitative light-induced fluorescence-digital imaging) and digital single-lens reflex camera of stained tooth surfaces might enable more efficient undertaking of large epidemiological studies and oral tests., Methods: Second-year dental hygiene students and registered dental hygienists 40 examiners were randomly selected, all of whom provided informed consent. Multiple examiners assessed plaque scoring indices from the two types of images of the same 30 patients. The images were observed on a computer monitor, and the area with dental plaque was assessed using the Patient Hygiene Performance Index. The intra-class correlation coefficient was determined to evaluate the agreement among examiners and test methods for dental plaque Q-ray cam and digital single-lens reflex camera measurements were compared using a Bland-Altman plot. The intra-class correlation coefficient was analyzed using SPSS version 24 software (SPSS Inc., Chicago, IL, USA), and the exploratory graph was analyzed with MedCalc., Results: Agreement among 40 examiners was excellent with an intra-class correlation coefficient of 0.990 when the Q-ray cam was used. The intra-class correlation coefficient for the registered dental hygienists group was 0.987 and that for the dental hygiene students group was 0.980., Conclusions: The study was high agreement in the assessment of Q-ray cam images and digital single-lens reflex camera images of stained dental plaque by multiple examiners, confirming that the two methods are validated both independently and with high mutual agreement., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A revolution in functional brain imaging techniques is in progress in the field of neurosciences. Optical imaging techniques, such as high-density diffuse optical tomography (HD-DOT), in which source-detector pairs of probes are placed on subjects' heads, provide better portability than conventional functional magnetic resonance imaging (fMRI) equipment. However, these techniques remain costly and can only acquire images at up to a few measurements per square centimetre, even when multiple detector probes are employed. In this study, we demonstrate functional brain imaging using a compact and affordable setup that employs nanosecond-order pulsed ordinary laser diodes and a time-extracted image sensor with superimposition capture of scattered components. Our technique can simply and easily attain a high density of measurement points without requiring probes to be attached, and can directly capture two-dimensional functional brain images. We have demonstrated brain activity imaging using a phantom that mimics the optical properties of an adult human head, and with a human subject, have measured cognitive brain activation while the subject is solving simple arithmetical tasks.

Multichannel (multicolor) imaging has become a powerful technique in biology research for performing in vivo neuronal calcium imaging, colocalization of fluorescent labels, non-invasive pH measurement, and other procedures. We describe a novel add-on approach for simultaneous multichannel optical microscopy based on simple wedge prisms. Our device requires no alignment and is simple, robust, user-friendly, and less expensive than current commercial instruments based on switchable filters or dual-view strategies. Point spread function measurements and simulations in Zemax indicate a reduction in resolution in the direction orthogonal to the wedge interface and in the axial direction, without introducing aberration. These effects depend on the objective utilized and are most significant near the periphery of the field of view. We tested a two-channel device on C. elegans neurons in vivo and demonstrated comparable signals to a conventional dual-view instrument. We also tested a four-channel device on fixed chick embryo Brainbow samples and identified individual neurons by their spectra without extensive image postprocessing. Therefore, we believe that this technology has the potential for broad use in microscopy.

Liquid biopsy with circulating tumor cells (CTCs) can aid in cancer detection at early stages and determine whether a certain treatment is effective or not. However, existing CTC techniques focus on one or two aspects of CTC management including sampling, enrichment, enumeration, and treatment selection. This paper reports an integrated capillary microchannel device that allows efficient capturing of CTCs with a wide microchannel, rapid enumeration of captured CTCs with whole slide cell imaging, and in situ drug testing with captured CTCs. Blood is drawn into the microchannel whose height is appropriate to the diameter of cancer cells, while its width is a thousand times larger than the diameter of cancer cells. The inner bottom surface of the microchannel is modified with long chain polymers that have cell adhesive ends to efficiently capture CTCs from blood. With this design, cells including CTCs are forced to move through the polymer coated microchannel, and the chance of cell adhesive ends interacting with specific antigens overexpressed on surfaces of cancer cells is significantly enhanced without a channel blockage issue. Captured CTCs are enumerated with a whole slide imaging platform via dual LED autofocusing technology then exposed to anti-cancer drugs, followed by live/dead assay and fluorescence imaging. Given its straightforward, easy and powerless operation, this device with whole slide imaging will be useful for cancer diagnosis, prognosis and point-of-care treatments.

Imaging technologies play a vital role in clinical oncology and have undergone massive growth over the past few decades. Research in the field of tumor imaging and biomedical diagnostics requires early detection of physiological alterations so as to provide curative treatment in real time. The objective of this review is to provide an insight about near infrared fluorescence (NIRF) and surface enhanced Raman scattering (SERS) imaging techniques that can be used to expand their capabilities for the early detection and diagnosis of cancer cells. Basic setup, principle and working of the instruments has been provided and common NIRF imaging agents as well as SERS tags are also discussed besides the analytical advantages/disadvantages of these techniques. This review can help researchers working in the field of molecular imaging to design cost effective fluorophores and SERS tags to overcome the limitations of both NIRF as well as SERS imaging technologies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Despite recent advances in cancer treatment, developing better therapeutic reagents remains an essential task for oncologists. To accurately characterize drug efficacy, 3D cell culture holds great promise as opposed to conventional 2D monolayer culture. Due to the advantages of cell manipulation in high-throughput, various microfluidic platforms have been developed for drug screening with 3D models. However, the dissemination of microfluidic technology is overall slow, and one missing part is fast and low-cost assay readout. In this work, we developed a microfluidic chip forming 1920 tumor spheres for drug testing, and the platform is supported by automatic image collection and cropping for analysis. Using conventional LIVE/DEAD staining as the ground truth of sphere viability, we trained a convolutional neural network to estimate sphere viability based on its bright-field image. The estimated sphere viability was highly correlated with the ground truth ( R -value > 0.84). In this manner, we precisely estimated drug efficacy of three chemotherapy drugs, doxorubicin, oxaliplatin, and irinotecan. We also cross-validated the trained networks of doxorubicin and oxaliplatin and found common bright-field morphological features indicating sphere viability. The discovery suggests the potential to train a generic network using some representative drugs and apply it to many different drugs in large-scale screening. The bright-field estimation of sphere viability saves LIVE/DEAD staining reagent cost and fluorescence imaging time. More importantly, the presented method allows viability estimation in a label-free and nondestructive manner. In short, with image processing and machine learning, the presented method provides a fast, low-cost, and label-free method to assess tumor sphere viability for large-scale drug screening in microfluidics.

Objective: To compare the AV Magnivisualizer with colposcopy to detect cervical intraepithelial neoplasia (CIN) using the Swede scoring system., Methods: Cross-sectional study conducted in a tertiary care hospital from May 2017 to March 2018. One hundred women with positive results at visual inspection with acetic acid underwent cervical inspection using the Magnivisualizer followed by a colposcope. Biopsies were taken if the Swede score was greater than 4. Pregnant women, women with an obvious cervical growth, acute cervicitis, or prior cervical surgery were excluded. Diagnostic accuracy of the Magnivisualizer and colposcope was calculated for high-grade lesions (CIN 2/CIN 2+) and agreement was compared between the two modalities., Results: The sensitivity, specificity, and positive and negative predictive values of the Magnivisualizer were 88.2%, 70.0%, 50.0%, and 94.6%, respectively, to detect high-grade lesions at a Swede score cutoff of 5 or more. The area under the curve for the Magnivisualizer was 0.80 (95% CI, 0.67-0.92), which was comparable with colposcopy (AUC 0.86; 95% CI, 0.76-0.96). There was very good agreement between the Magnivisualizer and colposcopy (κ=0.865, P<0.001) for high-grade lesions., Conclusion: The Magnivisualizer had high diagnostic accuracy to detect high-grade CIN in screen-positive women, which was comparable with colposcopy., (© 2019 International Federation of Gynecology and Obstetrics.)

Introduction: The incidence of malignant melanoma has been rising steadily over the past decades and Merkel cell carcinoma is a highly aggressive neuroendocrine skin tumor with a high mortality rate. Sentinel lymph node (SLN) biopsy is a recommended prognostic tool in primary cutaneous malignant melanomas of intermediate thickness and in all clinically node-negative Merkel cell carcinomas. The gold standard method for identification of SLNs is lymphoscintigraphy, which involves radioactive tracers. Indocyanine green-based near-infrared fluorescence imaging (NIRFI) has been also used for intraoperative SLN identification. We aim to evaluate the diagnostic sensitivity of the VisionSense VS3 NIRFI device for SNL identification. This device uses stereoscopic 3D high definition for both fluorescence and visible light imaging. Our hypothesis is that SLNs may be identified transcutaneously using fluorescent dye injections and NIRFI; therefore, obviating the need for lymphoscintigraphy in the future. METHODS AND ANALYSIS:: lymph node identification in skin malignancy using indocyanine green transcutaneously is a prospective diagnostic sensitivity study conducted at the Department of Plastic and Hand Surgery at the University Hospital Berne, Inselspital, Switzerland. The study aims at recruiting 93 patients (start date September 2017) to compare the accuracy of VisionSense VS3 camera at identifying SLNs transcutaneously with the current gold standard, lymphoscintigraphy. Moreover, a secondary objective is to determine if anatomical location of the SLN and patient factors (eg, body mass index, age) have an impact on the ability of VisionSense to detect SLNs when compared with the same gold standard., Trial Registration Number: ClinicalTrials.gov NCT03545334.

Multiphoton microscopy provides a suitable technique for imaging biological tissues with submicrometer resolution. Usually a Gaussian beam (GB) is used for illumination, leading to a reduced power efficiency in the multiphoton response and vignetting for a square-shaped imaging area. A flat-top beam (FTB) provides a uniform spatial intensity distribution that equalizes the probability of a multiphoton effect across the imaging area. We employ a customized widefield multiphoton microscope to compare the performance of a square-shaped FTB illumination with that based on using a GB, for both two-photon fluorescence (TPF) and second-harmonic generation (SHG) imaging. The variation in signal-to-noise ratio across TPF images of fluorescent dyes spans ∼5.6  dB for the GB and ∼1.2  dB for the FTB illumination, respectively. For the GB modality, TPF images of mouse colon and Convallaria root, and SHG images of chicken tendon and human breast biopsy tissue showcase ∼20  %   area that are not imaged due to either insufficient or lack of illumination. For quantitative analysis that depends on the illuminated area, this effect can potentially lead to inaccuracies. This work emphasizes the applicability of FTB illumination to multiphoton applications.

We report a new biofunctionalized nanoplatform based on hyaluronic acid-coated gold-nano-dot-decorated hollow carbon nanospheres (AuHCNs-HA) for microRNA imaging in living cells. Importantly, the HA-coated nanoplatform could be internalized into target cells via CD44 receptor-mediated endocytosis. It can be further applied for intracellular miR-21 imaging in CD44-positive colorectal cancer cells.

We propose a spectral contrast method to map the transmission images of surface plasmon resonance (SPR) in metallic nanostructures. Comparing the intensities between two neighboring wavelength bands near the SPR wavelength, the signal-to-noise ratio for biosensing applications obtained using the proposed method is found to be ten times higher than that obtained by conventional intensity analysis and 1.6 times better than that obtained by peak-wavelength fitting. The dynamic range and linearity of the refractive index are comparable to the peak-wavelength shift measurement. Based on the detection method, a spectral modulation system for the optical microscope is developed, combined with a gold-capped nanowire array, to measure the biointeractions in microfluidic devices. The experimental results show that the proposed method obtained multiple detections with a detection limit of 1.04 × 10 -5 refractive index units. Two types of analysis methods for SPR images are used to study the protein-antibody interactions. The region-of-interest analysis supports multiplexing detections in a compact microfluidic sensor. The effective pixel analysis eliminates low-response pixels and enhances the signal-to-noise ratios for sensitive label-free detection., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this report, an integrated optical platform based on spatial illumination together with laser speckle contrast technique was utilized to measure multiple parameters in live tissue including absorption, scattering, saturation, composition, metabolism, and blood flow. Measurements in three models of tissue injury including drug toxicity, artery occlusion, and acute hyperglycemia were used to test the efficacy of this system. With this hybrid apparatus, a series of structured light patterns at low and high spatial frequencies are projected onto the tissue surface and diffuse reflected light is captured by a CCD camera. A six position filter wheel, equipped with four bandpass filters centered at wavelengths of 650, 690, 800 and 880 nm is placed in front of the camera. Then, light patterns are blocked and a laser source at 650 nm illuminates the tissue while the diffusely reflected light is captured by the camera through the two remaining open holes in the wheel. In this manner, near-infrared (NIR) and laser speckle images are captured and stored together in the computer for off-line processing to reconstruct the tissue's properties. Spatial patterns are used to differentiate the effects of tissue scattering from those of absorption, allowing accurate quantification of tissue hemodynamics and morphology, while a coherent light source is used to study blood flow changes, a feature which cannot be measured with the NIR structured light. This combined configuration utilizes the strengths of each system in a complementary way, thus collecting a larger range of sample properties. In addition, once the flow and hemodynamics are measured, tissue oxygen metabolism can be calculated, a property which cannot be measured independently. Therefore, this merged platform can be considered a multiparameter wide-field imaging and spectroscopy modality. Overall, experiments demonstrate the capability of this spatially coregistered imaging setup to provide complementary, useful information of various tissue metrics in a simple and noncontact manner, making it attractive for use in a variety of biomedical applications., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Oral cancer is a growing health issue in low- and middle-income countries due to betel quid, tobacco, and alcohol use and in younger populations of middle- and high-income communities due to the prevalence of human papillomavirus. The described point-of-care, smartphone-based intraoral probe enables autofluorescence imaging and polarized white light imaging in a compact geometry through the use of a USB-connected camera module. The small size and flexible imaging head improves on previous intraoral probe designs and allows imaging the cheek pockets, tonsils, and base of tongue, the areas of greatest risk for both causes of oral cancer. Cloud-based remote specialist and convolutional neural network clinical diagnosis allow for both remote community and home use. The device is characterized and preliminary field-testing data are shared.

Background: Biomedical imaging devices that utilize the optical characteristics of hemoglobin (Hb) have become widespread. In the field of gastroenterology, there is a strong demand for devices that can apply this technique to surgical navigation. We aimed to introduce our novel multispectral device capable of intraoperatively performing quantitative imaging of the oxygen (O 2 ) saturation and Hb amount of tissues noninvasively and in real time, and to examine its application for deciding the appropriate anastomosis point after subtotal or total esophagectomy., Materials and Methods: A total of 39 patients with esophageal cancer were studied. Tissue O 2 saturation and Hb amount of the gastric tube just before esophagogastric anastomosis were evaluated using a multispectral tissue quantitative imaging device. The anastomosis point was decided depending on the quantitative values and patterns of both the tissue O 2 saturation and Hb amount., Results: The device can instantaneously and noninvasively quantify and visualize the tissue O 2 saturation and Hb amount using reflected light. The tissue Hb status could be classified into the following four types: good circulation type, congestion type, ischemia type, and mixed type of congestion and ischemia. Postoperative anastomotic failure occurred in 2 cases, and both were mixed cases., Conclusions: The method of quantitatively imaging the tissue O 2 saturation and Hb level in real time and noninvasively using a multispectral device allows instantaneous determination of the anastomosis and related organ conditions, thereby contributing to determining the appropriate treatment direction., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Near-infrared (NIR) nanoprobes with fluorescence "Turn-On" property are advantageous in cancer diagnosis but, to the best of our knowledge, "smart" nanoprobe that simultaneously targets both biotin receptor and carboxylesterase (CES) for HepG2 tumor-dual targeted imaging has not been reported. Methods : Using CBT-Cys click condensation reaction, we rationally designed a "smart" NIR fluorescence probe H 2 N-Cys(StBu)-Lys(Biotin)-Ser(Cy5.5)-CBT ( NIR-CBT ) and used it to facilely prepare the fluorescence-quenched nanoparticle NIR-CBT-NP . Results : In vitro results indicated that, after NIR-CBT-NP was incubated with CES for 6 h, its fluorescence was turned "On" by 69 folds. Cell experiments verified that NIR-CBT-NP was uptaken by HepG2 cells via biotin receptor-assisted endocytosis and its fluorescence was turned "On" by intracellular CES hydrolysis. Moreover, NIR-CBT-NP was successfully applied to image both biotin receptor- and CES-overexpressing HepG2 tumors. Conclusion : Fluorescence-quenched nanoparticle NIR-CBT-NP was facilely prepared to actively target biotin receptor-overexpressing HepG2 cancer cells and turn the fluorescence "On" by intracellular CES hydrolysis for tumor-dual targeted imaging. We anticipate that our fluorescence "Turn-On" nanoparticle could be applied for liver cancer diagnosis in clinic in the near future., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Metastasis is one of the most threatening aspects of cervical cancer. We developed a method to intraoperatively map the primary tumor, metastasis and metastatic sentinel lymph nodes (SLNs), providing real-time intraoperative guidance in cervical cancer. Methods : TMTP1, a tumor metastasis targeting peptide, was employed to modify the indocyanine green (ICG)-loaded poly (ethylene glycol)- poly (lactic-co-glycolic acid) (PEG-PLGA) micelles. The cervical cancer subcutaneous tumor model and lung metastasis model were established to determine the active targeting of ICG-loaded TMTP1-PEG-PLGA micelles (ITM) for the primary tumor and occult metastasis of cervical cancer. Human cervical cancer HeLa cells engineered by firefly luciferase were injected into the right hocks of BALB/c nude mice to develop the SLN metastasis model. The ITM and control ICG-loaded PEG-PLGA micelles (IM) were injected into the right hind footpads in the SLN metastasis model, and the migration and retention of micelles were recorded under near-infrared fluorescence. K14-HPV16 transgenic mice were also used to detect the image capability of ITM to target cancerous lesions. Results : ITM could actively target imaging of the primary tumor and cervical cancer metastasis. ITM quickly diffused from the injection site to SLNs along lymphatic capillaries and remained in the SLNs for 12 h. Moreover, ITM specifically accumulated in the tumor metastatic SLNs (T-SLNs), which could be successfully distinguished from normal SLNs (N-SLNs). Conclusion : ITM could achieve active targeting of the primary tumor, metastasis and T-SLNs, providing precise and real-time intraoperative guidance for cervical cancer., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Simultaneous imaging and manipulation of a genetically defined neuronal population can provide a causal link between its activity and function. Here, we designed a miniaturized microscope (or 'miniscope') that allows fluorescence imaging and optogenetic manipulation at the cellular level in freely behaving animals. This miniscope has an integrated optical connector that accepts any combination of external light sources, allowing flexibility in the choice of sensors and manipulators. Moreover, due to its simple structure and use of open source software, the miniscope is easy to build and modify. Using this miniscope, we demonstrate the optogenetic silencing of hippocampal CA1 neurons using two laser light sources-one stimulating a calcium sensor (i.e., jGCaAMP7c) and the other serving as an optogenetic silencer (i.e., Jaws). This new miniscope can contribute to efforts to determine causal relationships between neuronal network dynamics and animal behavior., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

The growing demand for tissue engineered vascular grafts (TEVG) motivates the development of optimized fabrication and monitoring procedures. Bioreactors which provide physiologically-relevant conditions are important for improving holistic TEVG properties and performance. Herein we describe a fiber-based intraluminal imaging system that allows for in situ assessment of vascular materials and re-cellularization processes inside a bioreactor by simultaneous and co-registered measurements of endogenous fluorescence lifetime and exogenous marker fluorescence intensity. The lumen of 6 vascular grafts (∼4 mm diameter) were scanned by reciprocally rotating a 41° angle polished multimode optical fiber inside a protective glass tube with outer diameter of 3 mm. Tubular bovine pericardium constructs were recellularized using enhanced Green Fluorescent Protein (eGFP) transfected cells in a custom bioreactor. The imaging system has resolved consistently the cellular autofluorescence from that of tissue matrix in situ based on the lifetime fluorescence properties of endogenous molecular species. The location of the re-cellularized area was validated by the eGFP emission. Current results demonstrate the potential of this system as a valuable tool in tissue engineering for in situ studies of cell-tissue interactions in cylindrical or other 3-dimensional structures.

The emission of Cherenkov photons from human and animal tissue can be observed during clinical x-ray or particle beam irradiation. However, imaging this weak emission with the necessary single-photon sensitivity in the clinical room is challenging because of milliwatt-level ambient room lighting and the presence of stray high-energy radiation. In this Letter, we demonstrate, to the best of our knowledge, the first Cherenkov imaging with a time-gated quanta image sensor employing a large single-photon avalanche diode (SPAD) array. Detecting single Cherenkov photons was possible with high photon avalanche gain, fast temporal gating, and moderately high ∼7% photon detection probability. Single-bit digitization and active SPAD quenching enabled stray x-ray noise suppression and photon-noise-limited imaging in a clinical environment. This type of imaging allows the knowledge of location, shape, and surface dose of the therapeutic beam radiotherapy with the stability of solid state-based detection.

Continuous and longitudinal imaging of cerebral blood flow (CBF) variations provide vital information to investigate pathophysiology and interventions for a variety of neurological and cerebral diseases. An innovative noncontact speckle contrast diffuse correlation tomography (scDCT) system was downscaled and adapted for noninvasive imaging of CBF distributions in rat brain through intact scalp and skull. Algorithms for 2D mapping and 3D image reconstruction of CBF distributions were developed and optimized. The continuous imaging capability of the system was shown by imaging global CBF increases during CO 2 inhalations and regional CBF decreases across two hemispheres during sequential unilateral and bilateral common carotid artery ligations. The longitudinal imaging capability was demonstrated by imaging CBF variations over a long recovery period of 14 days after an acute stroke. Compared to the 2D mapping method, the 3D imaging method reduces partial volume effects, but needs more computation time for image reconstruction. Results from this study generally agree with those reported in the literature using similar protocols to induce CBF changes in rats. The scDCT enables a relatively large penetration depth (up to ∼10 mm), which is sufficient for transcranial brain measurements in small animals and human neonates. Ultimately, we expect to provide a noninvasive noncontact cerebral imager for basic neuroscience research in small animal models and clinical applications in human neonates., (Copyright © 2019. Published by Elsevier Inc.)

The generation of physiologically relevant in-vitro models of biological barriers can play a key role in understanding human diseases and in the development of more predictive methods for assessing toxicity and drug or nutrient absorption. Here, we present an advanced cell culture system able to mimic the dynamic environment of biological barriers while monitoring cell behaviour through real-time impedance measurements and imaging. It consists of a fluidic device with an apical and a basal flow compartment separated by a semi-permeable membrane. The main features of the device are the integration of sensing through transepithelial electrical impedance (TEEI) measurements and transparent windows for optical monitoring within a dual flow system. Caco-2 cells were cultured in the TEEI bioreactor under both flow and static conditions. Although no differences in the expression of peripheral actin and occludin were visible, the cells in dynamic conditions developed higher impedance values at low frequencies, indicative of a higher paracellular electrical impedance with respect to the static cultures. TEEI measurements at high frequency also enabled monitoring monolayer formation, which can be correlated with the observation of an RC behaviour in the impedance spectra. In particular, the cells subject to flow showed accelerated barrier formation and increased vitality with respect to the static controls, again highlighting the importance of dynamic conditions for epithelial cells., (Copyright © 2019. Published by Elsevier B.V.)

Context.—: The rapid evolution of optical imaging modalities in recent years has opened the opportunity for ex vivo tissue imaging, which has significant implications for surgical pathology practice. These modalities have promising potential to be used as next-generation digital microscopy tools for examination of fresh tissue, with or without labeling with contrast agents., Objective.—: To review the literature regarding various types of ex vivo optical imaging platforms that can generate digital images for tissue recognition with potential for utilization in anatomic pathology clinical practices., Data Sources.—: Literature relevant to ex vivo tissue imaging obtained from the PubMed database., Conclusions.—: Ex vivo imaging of tissues can be performed by using various types of optical imaging techniques. These next-generation digital microscopy tools have a promising potential for utilization in surgical pathology practice.

Intelligent polymeric micelles with fluorescence imaging feature have been emerged as promising tools for theranostics. However, conventional fluorescent dyes are limited by short wavelength excitation, interference of tissue autofluorescence, limited imaging depth and quenched emission in aggregation state. Methods : We synthesized a novel mPEG-SS-Poly (AEMA- co -TBIS) (mPEATss) copolymer to develop multifunctional polymeric micelles with great AIE feature for cancer therapy and AIE active two-photon bioimaging. The stimuli-responsive behavior and AIE active two-photon cell and tissue imaging as well as in vitro and in vivo antitumor ability of DOX-loaded mPEATss were studied. Results : mPEATss micelles showed excellent AIE active two-photon cell imaging ability and deep tissue imaging ability. Antitumor drug DOX could be encapsulated to form a drug-loaded micellar system with a small diameter of 65 nm. The disassembly and charge-conversion of mPEATss micelles could be triggered by acidic environment, resulting in accelerated drug release and great antitumor efficacy. In vivo , ex vivo imaging and in vivo pharmacokinetic study demonstrated that mPEATss micelles could efficiently accumulate in tumor sites, which ensured ideal anticancer effect. Conclusions : This pH and redox dual responsive and AIE active two-photon imaging polymeric micelles would be a promising candidate for theranostics., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

We developed a gas-imaging system (sniff-cam) for gaseous ethanol (EtOH) with improved sensitivity. The sniff-cam was applied to measure the extremely low concentration distribution of breath EtOH without the consumption of alcohol, which is related to the activity of the oral or gut bacterial flora. A ring-type ultraviolet-light-emitting diode was mounted around a camera lens as an excitation light source, which enabled simultaneous excitation and imaging of the fluorescence. In the EtOH sniff-cam, a nicotinamide adenine dinucleotide (NAD)-dependent alcohol dehydrogenase (ADH) was used to catalyze the redox reaction between EtOH and the oxidized form of NAD (NAD + ). Upon application of gaseous EtOH to the ADH-immobilized mesh that was soaked in an NAD + solution and placed in front of the camera, NADH was produced through an ADH-mediated reaction. NADH expresses fluorescence at an emission wavelength of 490 nm and excitation wavelength of 340 nm. Thus, the concentration distribution of EtOH was visualized by measuring the distribution of the fluorescence light intensity from NADH on the ADH-immobilized mesh surface. First, a comparison of image analysis methods based on the red-green-blue color (RGB) images and the optimization of the buffer pH and NAD + solution concentration was performed. The new sniff-cam showed a 25-fold greater sensitivity and broader dynamic range (20.6-300000 ppb) in comparison to those of the previously fabricated sniff-cam. Finally, we measured the concentration distribution of breath EtOH without alcohol consumption using the improved sniff-cam and obtained a value of 116.2 ± 35.7 ppb ( n = 10).

Characterizing cell behavior is important to modern medical diagnoses as the changes of cell behavior are often indicators of huge diseases. In order to gain enough information about cells, developing novel methods of cell sorting and imaging is an important task. With development of micro-fabrication technologies, more advanced miniaturized devices are applied to cell research. Here, a portable and easy-to-use chip with high-density periodic micro-well array is designed and fabricated to capture target cells specifically. Combining with aptamer-silver conjugates and FAM functioned report probes, the sandwich assay was successfully applied for imaging cells. Any well of the chip is carefully designed to provide abundant information on single cells. Since there are 19,200 microwells in a single chip, more information is available. Compared to other cells, such as HEK-293, MCF-7, U 2 OS and Ramos cells, the sandwich assay shows high specificity towards target cell CCRF-CEM. What's more, the applications of the chip can be further expanded to other cells imaging if suitable aptamers were selected. This high-density micro-well array of aptamer-silver conjugates is hopeful to play an important role in medical diagnosis in the future., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Microrobotics extends the reach of human-controlled machines to submillimeter dimensions. We introduce a microrobot that relies on optoelectronic tweezers (OET) that is straightforward to manufacture, can take nearly any desirable shape or form, and can be programmed to carry out sophisticated, multiaxis operations. One particularly useful program is a serial combination of "load," "transport," and "deliver," which can be applied to manipulate a wide range of micrometer-dimension payloads. Importantly, microrobots programmed in this manner are much gentler on fragile mammalian cells than conventional OET techniques. The microrobotic system described here was demonstrated to be useful for single-cell isolation, clonal expansion, RNA sequencing, manipulation within enclosed systems, controlling cell-cell interactions, and isolating precious microtissues from heterogeneous mixtures. We propose that the optoelectronic microrobotic system, which can be implemented using a microscope and consumer-grade optical projector, will be useful for a wide range of applications in the life sciences and beyond., Competing Interests: The authors declare no conflict of interest.

Genetic code expansion (GCE) technology allows the specific incorporation of functionalized noncanonical amino acids (ncAAs) into proteins. Here, we investigated the Diels-Alder reaction between trans-cyclooct-2-ene (TCO)-modified ncAAs, and 22 known and novel 1,2,4,5-tetrazine-dye conjugates spanning the entire visible wavelength range. A hallmark of this reaction is its fluorogenicity - the tetrazine moiety can elicit substantial quenching of the dye. We discovered that photoinduced electron transfer (PET) from the excited dye to tetrazine is the main quenching mechanism in red-absorbing oxazine and rhodamine derivatives. Upon reaction with dienophiles quenching interactions are reduced resulting in a considerable increase in fluorescence intensity. Efficient and specific labeling of all tetrazine-dyes investigated permits super-resolution microscopy with high signal-to-noise ratio even at the single-molecule level. The different cell permeability of tetrazine-dyes can be used advantageously for specific intra- and extracellular labeling of proteins and highly sensitive fluorescence imaging experiments in fixed and living cells., Competing Interests: Competing interestsThe authors declare no competing interests.