Molecular fluorescence imaging is a commonly used method in various biomedical fields and is undergoing rapid translation toward clinical applications. Color images are commonly superimposed with fluorescence measurements to provide orientation, anatomical information, and molecular tissue properties in a single image. New adaptive methods that produce a more robust composite image than conventional lime green alpha blending are presented and demonstrated herein. Moreover, visualization through temporal changes is showcased as an alternative for real-time imaging systems.

We present the Toast++ open-source software environment for solving the forward and inverse problems in diffuse optical tomography (DOT). The software suite consists of a set of libraries to simulate near-infrared light propagation in highly scattering media with complex boundaries and heterogeneous internal parameter distribution, based on a finite-element solver. Steady-state, time- and frequency-domain data acquisition systems can be modeled. The forward solver is implemented in C++ and supports performance acceleration with parallelization for shared and distributed memory architectures, as well as graphics processing computation. Building on the numerical forward solver, Toast++ contains model-based iterative inverse solvers for reconstructing the volume distribution of absorption and scattering parameters from boundary measurements of light transmission. A range of regularization methods are provided, including the possibility of incorporating prior knowledge of internal structure. The user can link to the Toast++ libraries either directly to compile application programs for DOT, or make use of the included MATLAB and PYTHON bindings to generate script-based solutions. This approach allows rapid prototyping and provides a rich toolset in both environments for debugging, testing, and visualization.

Three-dimensional profiling and tracking by digital holography microscopy (DHM) provide label-free and quantitative analysis of the characteristics and dynamic processes of objects, since DHM can record real-time data for microscale objects and produce a single hologram containing all the information about their three-dimensional structures. Here, we have utilized DHM to visualize suspended microspheres and microfibers in three dimensions, and record the four-dimensional trajectories of free-swimming cells in the absence of mechanical focus adjustment. The displacement of microfibers due to interactions with cells in three spatial dimensions has been measured as a function of time at subsecond and micrometer levels in a direct and straightforward manner. It has thus been shown that DHM is a highly efficient and versatile means for quantitative tracking and analysis of cell motility.

Detecting and concentrating cancer cells in peripheral blood is of great importance for cancer diagnosis and prognosis. Optically induced dielectrophoresis (ODEP) can achieve high resolution and low optical intensities, and the electrode pattern can be dynamically changed by varied light patterns. By changing the projected light pattern, it is demonstrated to separate high-purity gastric cancer cell lines. Traditionally, the purity of cancer cell isolation by negative selection is 0.9% to 10%; by positive selection it is 50% to 62%. An ODEP technology is proposed to enhance the purity of cancer cell isolation to about 77%.

Monte Carlo modeling of light transport in multilayered tissue (MCML) is modified to incorporate objects of various shapes (sphere, ellipsoid, cylinder, or cuboid) with a refractive-index mismatched boundary. These geometries would be useful for modeling lymph nodes, tumors, blood vessels, capillaries, bones, the head, and other body parts. Mesh-based Monte Carlo (MMC) has also been used to compare the results from the MCML with embedded objects (MCML-EO). Our simulation assumes a realistic tissue model and can also handle the transmission/reflection at the object-tissue boundary due to the mismatch of the refractive index. Simulation of MCML-EO takes a few seconds, whereas MMC takes nearly an hour for the same geometry and optical properties. Contour plots of fluence distribution from MCML-EO and MMC correlate well. This study assists one to decide on the tool to use for modeling light propagation in biological tissue with objects of regular shapes embedded in it. For irregular inhomogeneity in the model (tissue), MMC has to be used. If the embedded objects (inhomogeneity) are of regular geometry (shapes), then MCML-EO is a better option, as simulations like Raman scattering, fluorescent imaging, and optical coherence tomography are currently possible only with MCML.

Quantitative evaluation of the potential radiation hazards of scanning light sources in medical optical devices is critical. Currently, point scanning light sources of continuous radiation are treated as pulsed sources, where the dwell time at each point is equal to the pulse duration. This study compares the photothermal effects from scanning light and pulsed sources using numerical calculation for scanning without restricting aperture and with various spot sizes. The calculation results show that the thermal damage threshold of scanning source not restricted by measurement aperture does not significantly differ from that of pulsed source. Temporal temperature response and size-dependent photothermal effect also confirm the similarity between scanning and pulsed sources.

We developed a Monte Carlo-based simulator of optical coherence tomography (OCT) imaging for turbid media with arbitrary spatial distributions. This simulator allows computation of both Class I diffusive reflectance due to ballistic and quasiballistic scattered photons and Class II diffusive reflectance due to multiple scattered photons. It was implemented using a tetrahedron-based mesh and importance sampling to significantly reduce computational time. Our simulation results were verified by comparing them with results from two previously validated OCT simulators for multilayered media. We present simulation results for OCT imaging of a sphere inside a background slab, which would not have been possible with earlier simulators. We also discuss three important aspects of our simulator: (1) resolution, (2) accuracy, and (3) computation time. Our simulator could be used to study important OCT phenomena and to design OCT systems with improved performance.

X-ray luminescence optical tomography (XLOT) is an emerging hybrid imaging modality in which x-ray excitable particles (phosphor particles) emit optical photons when stimulated with a collimated x-ray beam. XLOT can potentially combine the high sensitivity of optical imaging with the high spatial resolution of x-ray imaging. For reconstruction of XLOT data, we compared two reconstruction algorithms, conventional filtered backprojection (FBP) and a new algorithm, x-ray luminescence optical tomography with excitation priors (XLOT-EP), in which photon propagation is modeled with the diffusion equation and the x-ray beam positions are used as reconstruction priors. Numerical simulations based on dose calculations were used to validate the proposed XLOT imaging system and the reconstruction algorithms. Simulation results showed nanoparticle concentrations reconstructed with XLOT-EP are much less dependent on scan depth than those obtained with FBP. Measurements at just two orthogonal projections are sufficient for XLOT-EP to reconstruct an XLOT image for simple source distributions. The heterogeneity of x-ray energy deposition is included in the XLOT-EP reconstruction and improves the reconstruction accuracy, suggesting that there is a need to calculate the x-ray energy distribution for experimental XLOT imaging.

Quantitative measurement of dynamic responses of unstained living cells is of great importance in many applications ranging from investigation of fundamental cellular functions to drug discoveries. Conventional optical methods for label-free cell-based assay examine cellular structural changes proximal to sensor surfaces under external stimuli, but require dedicated nanostructure-patterned substrates for operation. Here, we present a quantitative imaging method, spectral-domain optical coherence phase microscopy (SD-OCPM), as a viable optical platform for label-free cell-based assay. The instrument is based on a low-coherence interferometric microscope that enables quantitative depth-resolved phase measurement of a transparent specimen with high phase stability. We demonstrate SD-OCPM measurement of dynamic responses of human breast cancer cells (MCF-7) to 2-picolinic acid (PA) and histamine.

Coronal tooth lesions, such as caries, enamel cracking, and composite resin restoration cavities, have been observed by optical coherence tomography (OCT). This pilot study was performed to verify whether OCT could reveal details of root canals filled with resin core build-up. A dual-cure, one-step, self-etch adhesive system-bonding agent (Clearfil Bond SE ONE, Kuraray Noritake Dental) and dual-cure resin composite core material (Clearfil DC Core Automix ONE, Kuraray Noritake Dental) were used according to the manufacturer’s instructions in root canals. OCT was performed at three stages of the core build-up: after the post space preparation, after bonding application, and after resin core fabrication. The cementum was removed in the cementum absent group and the root was left untreated in the cementum present group. Bubbles were observed in the resin cores and gaps formed between the resin core and dentin. In the cementum absent group, the internal structure of the root could be visualized clearly compared with the cementum present group. The root internal structure could be observed by OCT and the image became clearer when cementum was removed.

The application of time domain (TD) fluorescence lifetime multiplexing for the detection of fluorescent proteins (FPs) in whole animals, in the presence of a strong background tissue autofluorescence and excitation light leakage is discussed. Tissue autofluorescence (AF) exhibits a nonexponential temporal response, distinct from the mono-exponential decay of FPs. This allows a direct separation of FP fluorescence from AF using a dual basis function approach. We establish the detection limits of this approach using in vitro and in vivo measurements. We also demonstrate, using an experimental model of lymph node metastasis, that FP-AF lifetime multiplexing provides a greater than 30-fold improvement in contrast-to-background ratio compared with continuous wave data. In addition, we show that TD detection can simultaneously discriminate between up to three red shifted FPs placed under the skin of a nude mouse based on their distinct fluorescence lifetimes.

Retinal image quality assessment (IQA) is a crucial process for automated retinal image analysis systems to obtain an accurate and successful diagnosis of retinal diseases. Consequently, the first step in a good retinal image analysis system is measuring the quality of the input image. We present an approach for finding medically suitable retinal images for retinal diagnosis. We used a three-class grading system that consists of good, bad, and outlier classes. We created a retinal image quality dataset with a total of 216 consecutive images called the Diabetic Retinopathy Image Database. We identified the suitable images within the good images for automatic retinal image analysis systems using a novel method. Subsequently, we evaluated our retinal image suitability approach using the Digital Retinal Images for Vessel Extraction and Standard Diabetic Retinopathy Database Calibration level 1 public datasets. The results were measured through the F1 metric, which is a harmonic mean of precision and recall metrics. The highest F1 scores of the IQA tests were 99.60%, 96.50%, and 85.00% for good, bad, and outlier classes, respectively. Additionally, the accuracy of our suitable image detection approach was 98.08%. Our approach can be integrated into any automatic retinal analysis system with sufficient performance scores.

We propose a calibration-free photoacoustic (PA) method for transverse flow measurements. In this method, a pulsed periodically structured (i.e., grating patterned) optical beam is used to illuminate flowing absorptive particles in an optically scattering medium. The PA signal amplitudes measured over consecutive laser pulses carry an imprint of the illumination structure. The modulation frequency of the imprint is proportional to the component of the flow speed projected onto the normal axis of the striped illumination pattern. This method can tolerate high particle density, and is insensitive to the particle size, thus calibration-free. Bovine blood and microsphere phantoms were used to validate the proposed method. Blood flow in a mouse ear was measured in vivo as well.

We present the first experimental results of time-resolved diffuser-aided diffuse optical imaging (DADOI) method in this paper. A self-manufactured diffuser plate was inserted between the optode and the surface of a scattering medium. The diffuser was utilized to enhance the multiple scattering that destroys the image information for baseline measurement of turbid medium. Therefore, the abnormality can be detected with the modified optical density calculation. The time-domain DADOI method can provide better imaging contrast and simpler imaging than the conventional diffuse optical tomography measurement. Besides, it also reveals rich depth information with temporal responses. Therefore, the DADOI offers a great potential to detect the breast tumor and chemotherapy monitoring in clinical diagnosis.

A successful application of self-interference digital holographic microscopy in combination with a sample-rotation-based tomography module for three-dimensional (3-D) label-free quantitative live cell imaging with subcellular resolution is demonstrated. By means of implementation of a hollow optical fiber as the sample cuvette, the observation of living cells in different 3-D matrices is enabled. The fiber delivers a stable and accurate rotation of a cell or cell cluster, providing quantitative phase data for tomographic reconstruction of the 3-D refractive index distribution with an isotropic spatial resolution. We demonstrate that it is possible to clearly distinguish and quantitatively analyze several cells grouped in a “3-D cluster” as well as subcellular organelles like the nucleoli and local internal refractive index changes.

Hyperspectral imaging is a versatile tool that has recently been applied to a variety of biomedical applications, notably live-cell and whole-tissue signaling. Traditional hyperspectral imaging approaches filter the fluorescence emission over a broad wavelength range while exciting at a single band. However, these emission-scanning approaches have shown reduced sensitivity due to light attenuation from spectral filtering. Consequently, emission scanning has limited applicability for time-sensitive studies and photosensitive applications. In this work, we have developed an excitation-scanning hyperspectral imaging microscope that overcomes these limitations by providing high transmission with short acquisition times. This is achieved by filtering the fluorescence excitation rather than the emission. We tested the efficacy of the excitation-scanning microscope in a side-by-side comparison with emission scanning for detection of green fluorescent protein (GFP)-expressing endothelial cells in highly autofluorescent lung tissue. Excitation scanning provided higher signal-to-noise characteristics, as well as shorter acquisition times (300  ms/wavelength band with excitation scanning versus 3  s/wavelength band with emission scanning). Excitation scanning also provided higher delineation of nuclear and cell borders, and increased identification of GFP regions in highly autofluorescent tissue. These results demonstrate excitation scanning has utility in a wide range of time-dependent and photosensitive applications.

Cortical spreading depression (CSD) is a slowly propagating wave of depolarization of neurons and glia and has a less characterized vascular component. CSD is a commonly used phenomenon to test new methods of live brain imaging. Application of a blood pulsations imaging (BPI) technique to study of CSD induced with high-potassium solution in rat cortex allowed us to visualize for the first time the novel vascular component of a CSD wave. In our study, this wave component propagated in the limited part of the cortex along the bow-shaped trajectory in sharp contrast with concentric development of CSD measured by concurrently applied optical intrinsic signal (OIS) imaging technique. It was associated with a significant increase of the blood pulsations amplitude (BPA), started with a delay of 20 to 90 s comparing to signal measured with OIS, and propagated 40% faster than OIS signal. These findings suggest that the BPA and slower change of the cerebral blood volume are not directly related to each other even though both characterize the same vascular system. Our study indicates that the BPI technique could be used for characterization of the new pulsatile vascular component of CSDs in animal models of migraine, stroke, and brain trauma.

Intraoperative fluorescence molecular imaging based on targeted fluorescence agents is an emerging approach to improve surgical and endoscopic imaging and guidance. Short exposure times per frame and implementation at video rates are necessary to provide continuous feedback to the physician and avoid motion artifacts. However, fast imaging implementations also limit the sensitivity of fluorescence detection. To improve on detection sensitivity in video rate fluorescence imaging, we considered herein an optical flow technique applied to texture-rich color images. This allows the effective accumulation of fluorescence signals over longer, virtual exposure times. The proposed correction scheme is shown to improve signal-to-noise ratios both in phantom experiments and in vivo tissue imaging.

We demonstrate an extended-imaging-range anterior-segment optical coherence tomography (eAS-OCT) system for the biometric assessment of full AS in human eye. This newly developed eAS-OCT operating at 1340-nm wavelength band is simultaneously capable of an imaging speed of 120 kHz A-line scan rate, an axial resolution of 7.2 μm, and an extended imaging range of up to 16 mm in air. Imaging results from three healthy subjects and one subject with a narrow-angle demonstrate the instrument’s utility. With this system, it can provide anatomical dimensions of AS, including central corneal thickness, anterior chamber width, anterior chamber depth, crystalline lens vault, crystalline lens thickness, angle opening distance (AOD500/AOD750), and the area described by the trabecular-iris space (TISA500/TISA750) at 500/750  μm. We also use eAS-OCT to image and quantify dynamic functional changes of the AS in response to a light stimulus that induces physiological pupillary changes as well as accommodative efforts that induce lens changes. The results show that the described eAS-OCT is able to provide full anatomical biometry for AS and is useful for the studies where the dynamic response of AS compartment to certain stimulus is required.

In vivo optical imaging is widely used in preclinical studies. Recently, the application of optical imaging systems for preclinical visualization of gamma-emitting isotopes has become of interest since the evaluation of various organs relies on Tc^99m-based radiopharmaceuticals (RPs). In vitro radioluminescence of Tc^99m-based RPs, including pertechnetate, albumin macroaggregates, dimercaptosuccinic acid, phytate colloid, and ethylenediamine tetramethylene phosphonic acid, was studied with IVIS Spectrum CT™ optical imaging system. The distribution of phytate colloid was also studied in vivo with and without scintillating materials and the results were compared with those obtained with a conventional scintigraphy. The visible light emission appeared to be due to the radioluminescence of water and luminophores contained in RPs rather than from Cherenkov radiation. Weak air luminescence affected the background. The radioluminescence of fluids induced by Tc^99m-based tracers could be detected using charge-coupled device optical imaging systems. The radioluminescence intensity and its spectral distribution depend on the surrounding fluid and known luminophores present. Thus, in some cases the in vivo optical imaging is possible but the use of scintillator, e.g., borosilicate glass or bismuth germanate, is preferred.

The optical activity of glucose molecules is important for diagnosing and monitoring blood glucose of diabetes. In order to accurately detect the diabetes at an early stage, there is an urgent need to develop innovative detection methods. By use of Mueller matrix decomposition (MMD), we have studied the optical activity of glucose molecules in three types of turbid media—polystyrene (PST) sphere suspension, chicken blood, and the vein blood of diabetic patients. The experimental setup to obtain the Mueller matrix in the forward detection geometry has been used. The experimental results show that the rotation angle has linear relationship with the concentration of the glucose when the scattering coefficient of the PST sphere suspension remains unchanged, whereas the scattering effect enlarges the rotation angle. Furthermore, optical rotation abides by Drude’s dispersion equation. The decomposition method has also been found useful applications in quantifying the optical rotations of blood glucose in diabetic patients. The diabetic severity status can be distinguished with the rotation angle of glucose and also is in accordance with the clinical diagnosis. Thus, the method of MMD has promising applications in diabetic diagnosis and monitoring.

An advantage of fluorescence methods over other imaging modalities is the ability to concurrently resolve multiple moieties using fluorochromes emitting at different spectral regions. Simultaneous imaging of spectrally separated agents is helpful in interrogating multiple functions or establishing internal controls for accurate measurements. Herein, we investigated multimoiety imaging in the context of a limited-projection-angle hybrid fluorescence molecular tomography (FMT), and x-ray computed tomography implementation and the further registration with positron emission tomography (PET) data. Multichannel FMT systems may image fluorescent probes of varying distribution patterns. Therefore, it is possible that different channels may require different use of priors and regularization parameters. We examined the performance of automatically estimating regularization factors implementing priors, using data-driven regularization specific for limited-projection-angle schemes. We were particularly interested in identifying the implementation variations between hybrid-FMT channels due to probe distribution variation. For this reason, initial validation of the data-driven algorithm on a phantom was followed by imaging different agent distributions in animals, assuming superficial and deep seated activity. We further demonstrate the benefits of combining hybrid FMT with PET to gain multiple readings on the molecular composition of disease.

Protein-protein interactions in cells are often studied using fluorescence resonance energy transfer (FRET) phenomenon by fluorescence lifetime imaging microscopy (FLIM). Here, we demonstrate approaches to the quantitative analysis of FRET in cell population in a case complicated by a highly heterogeneous donor expression, multiexponential donor lifetime, large contribution of cell autofluorescence, and significant presence of unquenched donor molecules that do not interact with the acceptor due to low affinity of donor-acceptor binding. We applied a multifrequency phasor plot to visualize FRET FLIM data, developed a method for lifetime background correction, and performed a detailed time-resolved analysis using a biexponential model. These approaches were applied to study the interaction between the Toll Interleukin-1 receptor (TIR) domain of Toll-like receptor 4 (TLR4) and the decoy peptide 4BB. TLR4 was fused to Cerulean fluorescent protein (Cer) and 4BB peptide was labeled with Bodipy TMRX (BTX). Phasor displays for multifrequency FLIM data are presented. The analytical procedure for lifetime background correction is described and the effect of correction on FLIM data is demonstrated. The absolute FRET efficiency was determined based on the phasor plot display and multifrequency FLIM data analysis. The binding affinity between TLR4-Cer (donor) and decoy peptide 4BB-BTX (acceptor) was estimated in a heterogeneous HeLa cell population.

Owing to the high degree of scattering of light through tissues, the ill-posedness of fluorescence molecular tomography (FMT) inverse problem causes relatively low spatial resolution in the reconstruction results. Unlike L2 regularization, L1 regularization can preserve the details and reduce the noise effectively. Reconstruction is obtained through a restarted L1 regularization-based nonlinear conjugate gradient (re-L1-NCG) algorithm, which has been proven to be able to increase the computational speed with low memory consumption. The algorithm consists of inner and outer iterations. In the inner iteration, L1-NCG is used to obtain the L1-regularized results. In the outer iteration, the restarted strategy is used to increase the convergence speed of L1-NCG. To demonstrate the performance of re-L1-NCG in terms of spatial resolution, simulation and physical phantom studies with fluorescent targets located with different edge-to-edge distances were carried out. The reconstruction results show that the re-L1-NCG algorithm has the ability to resolve targets with an edge-to-edge distance of 0.1 cm at a depth of 1.5 cm, which is a significant improvement for FMT.

Acne vulgaris as an inflammatory disease, with an excessive production of subdermal fat, modifies the dynamics of the bloodstream, and consequently temperature, on the affected skin zone. A high percentage of this heat interchange is manifested as electromagnetic radiation with far-infrared wavelengths, which can be captured through a thermal imaging camera. A technique based on thermal image analysis for efficiency assessment in acne vulgaris is described. The procedure is based on computing statistical parameters of thermal images captured from the affected skin zone being attended by an acne treatment. The proposed technique was used to determine the skin thermal behavior according to acne severity levels in different acne treatment stages. Infrared images of acne skin zones on eight patients, diagnosed with acne vulgaris and attended by one specific acne treatment, were weekly registered during 11 weeks. The infrared images were captured until no more improvement in affected zones was detected. The obtained results suggest a direct relationship between the used statistical parameters, particularly first- and second-order statistics, and the acne vulgaris severity level on the affected zones.

Multispectral Mueller matrix imaging was performed over a spectral range from 470 to 632 nm on 4-μm unstained gastric tissue sections. A complete set of polarization parameters was derived. The combination of linear depolarization and linear retardance yields the highest accuracy in sample classification. When the depolarization of linearly polarized light due to scattering is independent of the orientation angle of the incident linear polarization vector, the derivation of linear polarization properties will require only 3×3 Mueller matrix, which would significantly reduce the complexity of the polarimetry imaging system. When additional parameters are needed to complement the two linear polarization parameters, retardance, circular depolarization, and depolarization can be included in classification in the order of preference. However, these additional parameters would require the measurement of 4×4 Mueller matrix. In addition, it appears that wavelength is not a critical factor in terms of classification accuracy for thin tissue sections in this study.

Our previous work demonstrated that an optical coherence tomography (OCT) technique was able to characterize the whole blood coagulation process. The 1/e light penetration depth (d _1/e ) derived from the profiles of reflectance versus depth was developed for detecting the whole blood coagulation process in static state. To consider the effect of blood flow, in the present study, d 1/e versus time from the coagulating porcine blood circulated in a mock flow loop with various steady laminar flows at mean flow speed in the range from 5 to 25  mm/s . The variation of d _1/e was used to represent the change of blood properties during coagulation in different hematocrits (HCT) ranging from 25% to 55%, velocities from 5 to 25  mm/s , and tubing sizes from 0.9 to 2 mm. The results showed that there were positive correlations between coagulation time (t _c ) and HCT, velocity, and tubing size, respectively. In addition, the coagulation rate (S _r ) was decreased with the increase of HCT, velocity, and tubing size. This study testified that HCT, flow velocity, and tubing size were substantial factors affecting the backscattering properties during flowing blood coagulation. Furthermore, OCT has the potential to represent the process of flowing blood coagulation with proper parameters.

The advantages of using a telecentric imaging system in digital holographic microscopy (DHM) to study biological specimens are highlighted. To this end, the performances of nontelecentric DHM and telecentric DHM are evaluated from the quantitative phase imaging (QPI) point of view. The evaluated stability of the microscope allows single-shot QPI in DHM by using telecentric imaging systems. Quantitative phase maps of a section of the head of the drosophila melanogaster fly and of red blood cells are obtained via single-shot DHM with no numerical postprocessing. With these maps we show that the use of telecentric DHM provides larger field of view for a given magnification and permits more accurate QPI measurements with less number of computational operations.

Ischemia-reperfusion injuries are a critical determinant of lung transplantation success. The endogenous production of carbon monoxide (CO) is triggered by ischemia-reperfusion injuries. Our aim was, therefore, to assess the feasibility of exhaled CO measurements during the ex vivo evaluation of lungs submitted to ischemia-reperfusion injuries. Five pigs were euthanized and their lungs removed after pneumoplegia. After cold storage (30 min, 4°C), the lungs were connected to an extracorporeal membrane oxygenation circuit, slowly warmed-up, and ventilated. At the end of a 45-min steady state, CO measurements were performed by optical-feedback cavity-enhanced absorption spectroscopy, a specific laser-based technique for noninvasive and real-time low gas concentration measurements. Exhaled CO concentration from isolated lungs reached 0.45±0.19  ppmv and was above CO concentration in ambient air and in medical gas. CO variations peaked during the expiratory phase. Changes in CO concentration in ambient air did not alter CO concentrations in isolated lungs. Exhaled CO level was also found to be uncorrelated to heme oxygenase (HO-1) gene expression. These results confirm the feasibility of accurate and real-time CO measurement in isolated lungs. The presented technology could help establishing the exhaled CO concentration as a biomarker of ischemia-reperfusion injury in ex vivo lung perfusion.

A biocompatible fiber-optic pH sensor based on a unique double-pass Mach–Zehnder interferometer is proposed. pH responsive poly(2-hydroxyethyl methacrylate-co-2-(dimethylamino)ethyl methacrylate) hydrogel coating on the fiber swells/deswells in response to local pH, leading to refractive index changes that manifest as shifting of interference dips in the optical spectrum. The pH sensor is tested in spiked phosphate buffer saline and demonstrates high sensitivity of 1.71  nm/pH , pH 0.004 limit of detection with good responsiveness, repeatability, and stability. The proposed sensor has been successfully applied in monitoring the media pH in cell culture experiments to investigate the relationship between pH and cancer cell growth.

The feasibility and utility of wearable 24-h multimodality neuromonitoring during daily activities are demonstrated. We have developed a fourth-generation ambulatory near infrared spectroscopy device, namely NINscan 4. NINscan 4 enables recording of brain function (via cerebral hemodynamics), systemic hemodynamics, electrocardiography, and actigraphy simultaneously and continuously for up to 24 h at 250-Hz sampling rate, during (and with minor restriction to) daily activities. We present initial 24-h human subject test results, with example analysis including (1) comparison of cerebral perfusion and oxygenation changes during wakefulness and sleep over a 24-h period and (2) capturing of hemodynamic changes prior, during and after sudden waken up in the night during sleep. These results demonstrate the first ambulatory 24-h cerebral and systemic hemodynamics monitoring, and its unique advantages including long-term data collection and analysis capability, ability to catch unpredictable transient events during activities of daily living, as well as coregistered multimodality analysis capabilities. These results also demonstrate that NINscan 4’s motion artifact at 1-g head movement is smaller than physiological hemodynamic fluctuations during motionless sleep. The broader potential of this technology is also discussed.

We present a time-resolved photographic analysis of the pulse-to-pulse interaction. In particular, we studied the influence of the cavitation bubble induced by a fs-pulse on the optical focusing of the consecutive pulse and its cavitation bubble dynamics in dependence on temporal pulse separation in water. As a first result, by decreasing the temporal separation of laser pulses, there is a diminishment of the laser-induced optical breakdown (LIOB) efficiency in terms of energy conversion, caused by disturbed focusing into persisting gas bubbles at the focal volume. A LIOB at the focal spot is finally suppressed by impinging the expanding or collapsing cavitation bubble of the preceding pulse. These results could be additionally confirmed in porcine gelatin solution with various concentrations. Hence, the interaction between the laser and transparent ophthalmic tissue may be accompanied by a raised central laser energy transmission, which could be observed in case of a temporal pulse overlap. In conclusion, our experimental results are of particular importance for the optimization of the prospective ophthalmic surgical process with future generation fs-lasers.

Laser phototherapy (LPT) is widely used in clinical practice to accelerate healing. Although the use of LPT has advantages, the molecular mechanisms involved in the process of accelerated healing and the safety concerns associated with LPT are still poorly understood. We investigated the physiological effects of LPT irradiation on the production and accumulation of reactive oxygen species (ROS), genomic instability, and deoxyribose nucleic acid (DNA) damage in human epithelial cells. In contrast to a high energy density (20  J/cm ² ), laser administered at a low energy density (4  J/cm ² ) resulted in the accumulation of ROS. Interestingly, 4  J/cm ² of LPT did not induce DNA damage, genomic instability, or nuclear influx of the BRCA1 DNA damage repair protein, a known genome protective molecule that actively participates in DNA repair. Our results suggest that administration of low energy densities of LPT induces the accumulation of safe levels of ROS, which may explain the accelerated healing results observed in patients. These findings indicate that epithelial cells have an endowed molecular circuitry that responds to LPT by physiologically inducing accumulation of ROS, which triggers accelerated healing. Importantly, our results suggest that low energy densities of LPT can serve as a safe therapy to accelerate epithelial healing.

This article [J. Biomed. Opt.. 19, (2 ), 027007 (2014)] was originally published online on 18 February 2014 with an error in the abstract. The values for highest sensitivity and specificity were stated to be 91% and 81%, respectively, but these were based on an earlier data analysis. The correct values were reported in the body of the paper. The values in the abstract have been corrected to align with those reported in the body of the paper, which are 89% for sensitivity and 86% for specificity. This article was corrected online on 14 March 2014.

Microscopes are tools invented by humans to study specimens at spatial and temporal resolution that exceed the human eye. Today the researcher has a wide range of microscopes to employ in order to answer carefully poised experimental questions. The key is to match the research questions with the appropriate microscope. In order to optimize the success it is necessary to understand the physical principles of the microscope, how to properly align the instrument, and how to correctly use the instrument. The object of the investigation or measurement is also a major consideration as well as the preparation of the specimen, the interaction of the specimen, and the radiation from the microscope. Often the words “quantitative microscopy” appear in publications; but all measurements contain errors, and the critical investigator must be able to understand the sources of error in a measurement and, if possible, mitigate the magnitude of the errors. Validation of microscopic measurements across disparate types of microscopes is a useful approach. The critical elements in optical microscopy include: optical aberrations, sampling errors, nonphysiological specimen preparation, such as over expression of genetic fluorescent proteins, the deleterious effects of the light source on biological specimens, nonlinear effects in the detector and amplifiers, and artifacts of image analysis and image interpretation.

Haishan Zeng, 272 pages, ISBN: 13:978-1-4200-8346-0, CRC Press, New York (2013) $99.95, hardcover. Reviewed by Lingyan Shi, research assistant and PhD candidate, Department of Biomedical Engineering, The City College of the City University of New York Diagnostic Endoscopy, edited by Haishan Zeng, is a new book recently published by CRC Press. This book is clearly organized and incorporates both well-established and new theories and principles of endoscopy that are used in laboratory and clinical practice. It will help enlighten the medical community to unlock the value of applying spectroscopy to medicine, and to educate undergraduate and graduate students in the field of biomedical optics.