A dual-modality birefringence/phase imaging system is presented. The system features a crystal retarder that provides polarization mixing and generates two interferometric carrier waves in a single signal spectrum. The retardation and orientation of sample birefringence can then be measured simultaneously based on spectral multiplexing interferometry. Further, with the addition of a Nomarski prism, the same setup can be used for quantitative differential interference contrast (DIC) imaging. Sample phase can then be obtained with two-dimensional integration. In addition, birefringence-induced phase error can be corrected using the birefringence data. This dual-modality approach is analyzed theoretically with Jones calculus and validated experimentally with malaria-infected red blood cells. The system generates not only corrected DIC and phase images, but a birefringence map that highlights the distribution of hemozoin crystals.

Quantifying tissue biomechanical properties can assist in detection of abnormalities and monitoring disease progression and/or response to a therapy. Optical coherence elastography (OCE) has emerged as a promising technique for noninvasively characterizing tissue biomechanical properties. Several mechanical loading techniques have been proposed to induce static or transient deformations in tissues, but each has its own areas of applications and limitations. This study demonstrates the combination of Lorentz force excitation and phase-sensitive OCE at ∼1.5  million A-lines per second to quantify the elasticity of tissue by directly imaging Lorentz force-induced elastic waves. This method of tissue excitation opens the possibility of a wide range of investigations using tissue biocurrents and conductivity for biomechanical analysis.

Ocular angiogenesis is a blinding complication of age-related macular degeneration and other retinal vascular diseases. Clinical imaging approaches to detect inflammation prior to the onset of neovascularization in these diseases may enable early detection and timely therapeutic intervention. We demonstrate the feasibility of a previously developed cyclooxygenase-2 (COX-2) targeted molecular imaging probe, fluorocoxib A, for imaging retinal inflammation in a mouse model of laser-induced choroidal neovascularization. This imaging probe exhibited focal accumulation within laser-induced neovascular lesions, with minimal detection in proximal healthy tissue. The selectivity of the probe for COX-2 was validated in vitro and by in vivo retinal imaging with nontargeted 5-carboxy-X-rhodamine dye, and by blockade of the COX-2 active site with nonfluorescent celecoxib prior to injection of fluorocoxib A. Fluorocoxib A can be utilized for imaging COX-2 expression in vivo for further validation as an imaging biomarker in retinal diseases.

This study demonstrates the feasibility of using the Rayleigh wave model (RWM) in combination with optical coherence elastography (OCE) technique to assess the viscoelasticity of soft tissues. Dispersion curves calculated from the spectral decomposition of OCE-measured air-pulse induced elastic waves were used to quantify the viscoelasticity of samples using the RWM. Validation studies were first conducted on 10% gelatin phantoms with different concentrations of oil. The results showed that the oil increased the viscosity of the gelatin phantom samples. This method was then used to quantify the viscoelasticity of chicken liver. The Young’s modulus of the chicken liver tissues was estimated as E=2.04±0.88  kPa with a shear viscosity η=1.20±0.13  Pa s. The analytical solution of the RWM correlated very well with the OCE-measured phased velocities (R²=0.96±0.04). The results show that the combination of the RWM and OCE is a promising method for noninvasively quantifying the biomechanical properties of soft tissues and may be a useful tool for detecting disease.

We use a quantitative phase imaging technique, defocusing microscopy (DM), to measure morphological, chemical, and mechanical parameters of individual red blood cells (RBCs) immersed in solutions with different osmolalities. We monitor the RBCs’ radius, volume, surface area, sphericity index, and hemoglobin content and concentration. The complete shape of cells is recovered and the effects of their adhesion to the glass substrate are observed. Finally, membrane fluctuation measurements give us information about the cells deformability.

An image reconstruction regularization approach for magnetic resonance imaging-guided near-infrared spectral tomography has been developed to improve quantification of total hemoglobin (HbT) and water. By combining prior information from dynamic contrast enhanced (DCE) and diffusion weighted (DW) MR images, the absolute bias errors of HbT and water in the tumor were reduced by 22% and 18%, 21% and 6%, and 10% and 11%, compared to that in the no-prior, DCE- or DW-guided reconstructed images in three-dimensional simulations, respectively. In addition, the apparent contrast values of HbT and water were increased in patient image reconstruction from 1.4 and 1.4 (DCE) or 1.8 and 1.4 (DW) to 4.6 and 1.6.

This study aimed to perform a systematic review to assess the effectiveness of antimicrobial photodynamic therapy (aPDT) in the reduction of microorganisms in deep carious lesions. An electronic search was conducted in Pubmed, Web of Science, Scopus, Lilacs, and Cochrane Library, followed by a manual search. The MeSH terms, MeSH synonyms, related terms, and free terms were used in the search. As eligibility criteria, only clinical studies were included. Initially, 227 articles were identified in the electronic search, and 152 studies remained after analysis and exclusion of the duplicated studies; 6 remained after application of the eligibility criteria; and 3 additional studies were found in the manual search. After access to the full articles, three were excluded, leaving six for evaluation by the criteria of the Cochrane Collaboration’s tool for assessing risk of bias. Of these, five had some risk of punctuated bias. All results from the selected studies showed a significant reduction of microorganisms in deep carious lesions for both primary and permanent teeth. The meta-analysis demonstrated a significant reduction in microorganism counts in all analyses (p<0.00001). Based on these findings, there is scientific evidence emphasizing the effectiveness of aPDT in reducing microorganisms in deep carious lesions.

This PDF file contains the editorial “Special Section Guest Editorial:Clinical near-infrared spectroscopy and imaging” for JBO Vol. 21 Issue 09

Near-infrared spectroscopy (NIRS) has been used to measure reactive hyperemia following a vascular occlusion. However, the procedures and methods of analysis used have varied. The purpose of the present study is to identify reproducible methods for measuring reactive hyperemia using HbO₂ NIRS signals in the calf and foot. Healthy participants (10 male, 10 female) aged 19 to 28 years performed one of two tests: reproducibility trials or elevation protocol (30 and 60 cm limb elevation above the heart). The time to 50% reperfusion (T_1/2) and the second (R_2q) quartile rates of reperfusion were found to be the most reproducible parameters (coefficient of variation=7.12 to 14.1%). The time to 95% reperfusion (T₉₅) was 12.7% more reproducible on average than the previously reported parameter of time to peak hyperemia. Measures of reperfusion time and rate slowed with increasing limb elevation. Correlations were identified between the calf and foot in the measurements of R_2q (R²=0.713, p=0.021), T_1/2 (R²=0.673, p=0.033), and T₉₅ (R²=0.792, p=0.006). Half and 95% recovery times and second and third quartile rates expressed good reproducibility and sensitivity to change with reduced perfusion pressure. NIRS measures of reactive hyperemia have the potential to evaluate microvascular perfusion in clinical populations.

In traditional brain–computer interface (BCI) studies, binary communication systems have generally been implemented using two mental tasks arbitrarily assigned to “yes” or “no” intentions (e.g., mental arithmetic calculation for “yes”). A recent pilot study performed with one paralyzed patient showed the possibility of a more intuitive paradigm for binary BCI communications, in which the patient’s internal yes/no intentions were directly decoded from functional near-infrared spectroscopy (fNIRS). We investigated whether such an “fNIRS-based direct intention decoding” paradigm can be reliably used for practical BCI communications. Eight healthy subjects participated in this study, and each participant was administered 70 disjunctive questions. Brain hemodynamic responses were recorded using a multichannel fNIRS device, while the participants were internally expressing “yes” or “no” intentions to each question. Different feature types, feature numbers, and time window sizes were tested to investigate optimal conditions for classifying the internal binary intentions. About 75% of the answers were correctly classified when the individual best feature set was employed (75.89% ±1.39 and 74.08% ±2.87 for oxygenated and deoxygenated hemoglobin responses, respectively), which was significantly higher than a random chance level (68.57% for p<0.001). The kurtosis feature showed the highest mean classification accuracy among all feature types. The grand-averaged hemodynamic responses showed that wide brain regions are associated with the processing of binary implicit intentions. Our experimental results demonstrated that direct decoding of internal binary intention has the potential to be used for implementing more intuitive and user-friendly communication systems for patients with motor disabilities.

Wound management is a challenging and costly problem that is growing in importance as people are living longer. Instrumental methods are increasingly being relied upon to provide objective measures of wound assessment to help guide management. Technologies that employ near-infrared (NIR) light form a prominent contingent among the existing and emerging technologies. We review some of these technologies. Some are already established, such as indocyanine green fluorescence angiography, while we also speculate on others that have the potential to be clinically relevant to wound monitoring and assessment. These various NIR-based technologies address clinical wound management needs along the entire healing trajectory of a wound.

¹⁸F-fluorodeoxyglucose positron emission tomography combined with computed tomography (FDG-PET/CT) is widely used as a standard method for evaluating human brown adipose tissue (BAT), a recognized therapeutic target of obesity. However, a longitudinal BAT study using FDG-PET/CT is lacking owing to limitations of the method. Near-infrared time-resolved spectroscopy (NIR_TRS) is a technique for evaluating human BAT density noninvasively. This study aimed to test whether NIR_TRS could detect changes in BAT density during or after long-term intervention. First, using FDG-PET/CT, we confirmed a significant increase (+48.8%, P<0.05) in BAT activity in the supraclavicular region after 6-week treatment with thermogenic capsaicin analogs, capsinoids. Next, 20 volunteers were administered either capsinoids or placebo daily for 8 weeks in a double-blind design, and BAT density was measured using NIR_TRS every 2 weeks during the 8-week treatment period and an 8-week period after stopping treatment. Consistent with FDG-PET/CT results, NIR_TRS successfully detected an increase in BAT density during the 8-week treatment (+46.4%, P<0.05), and a decrease in the 8-week follow-up period (−12.5%, P=0.07), only in the capsinoid-treated, but not the placebo, group. Thus, NIR_TRS can be applied for quantitative assessment of BAT in longitudinal intervention studies in humans.

Global tissue perfusion and oxygenation are important indicators of physiologic function in humans. The monitoring of splanchnic oximetry through the use of near-infrared spectroscopy (NIRS) is an emerging method used to assess tissue oxygenation status. Splanchnic tissue oxygenation (SrSO₂) is thought to be potentially of high value in critically ill patients because gastrointestinal organs can often be the first to suffer ischemic injury. During conditions of hypovolemia, cardiac dysfunction, or decreased oxygen-carrying capacity, blood flow is diverted toward vital organs, such as the brain and the heart at the expense of the splanchnic circulation. While monitoring SrSO₂ has great potential benefit, there are limitations to the technology and techniques. SrSO₂ has been found to have a relatively high degree of variability that can potentially make it difficult to interpret. In addition, because splanchnic organs only lie near the skin surface in children and infants, and energy from currently available sensors only penetrates a few centimeters deep, it can be difficult to use clinically in a noninvasive manner in adults. Research thus far is showing that splanchnic oximetry holds great promise in the ability to monitor patient oxygenation status and detect disease states in humans, especially in pediatric populations.

Near-infrared spectroscopy (NIRS) measurements of cytochrome-c-oxidase (CCO) have the potential to yield crucial information about cerebral metabolism at the patient bedside. Developments in instrumentation and the analytical methods used to resolve changes in CCO have led to many clinical applications of the measurement since its first demonstration in 1977 by Jöbsis. There is a substantial literature of work on measures of CCO in animal and in vitro studies; however, this review focuses on translational studies. Almost 40 years from the advent of the first measurement of CCO using NIRS, this signal continues to hold significant interest in our understanding of the human brain in health and disease. We discuss methodologies for obtaining NIRS measurements of CCO in the clinic and review studies in neonates and adults.

Pulse rate variability (PRV) can be extracted from functional near-infrared spectroscopy (fNIRS) (PRV_NIRS) and photoplethysmography (PPG) (PRV_PPG) signals. The present study compared the accuracy of simultaneously acquired PRV_NIRS and PRV_PPG, and evaluated their different characterizations of the sympathetic (SNS) and parasympathetic (PSNS) autonomous nervous system activity. Ten healthy subjects were recorded during resting-state (RS) and respiratory challenges in two temperature conditions, i.e., room temperature (23°C) and cold temperature (4°C). PRV_NIRS was recorded based on fNIRS measurement on the head, whereas PRV_PPG was determined based on PPG measured at the finger. Accuracy between PRV_NIRS and PRV_PPG, as assessed by cross-covariance and cross-sample entropy, demonstrated a high degree of correlation (r<0.9), which was significantly reduced by respiration and cold temperature. Characterization of SNS and PSNS using frequency-domain, time-domain, and nonlinear methods showed that PRV_NIRS provided significantly better information on increasing PSNS activity in response to respiration and cold temperature than PRV_PPG. The findings show that PRV_NIRS may outperform PRV_PPG under conditions in which respiration and temperature changes are present, and may, therefore, be advantageous in research and clinical settings, especially if characterization of the autonomous nervous system is desired.

Fluorescence imaging has been considered for over a half-century as a modality that could assist surgical guidance and visualization. The administration of fluorescent molecules with sensitivity to disease biomarkers and their imaging using a fluorescence camera can outline pathophysiological parameters of tissue invisible to the human eye during operation. The advent of fluorescent agents that target specific cellular responses and molecular pathways of disease has facilitated the intraoperative identification of cancer with improved sensitivity and specificity over nonspecific fluorescent dyes that only outline the vascular system and enhanced permeability effects. With these new abilities come unique requirements for developing phantoms to calibrate imaging systems and algorithms. We briefly review herein progress with fluorescence phantoms employed to validate fluorescence imaging systems and results. We identify current limitations and discuss the level of phantom complexity that may be required for developing a universal strategy for fluorescence imaging calibration. Finally, we present a phantom design that could be used as a tool for interlaboratory system performance evaluation.

The recent developments in time-domain diffuse optics that rely on physical concepts (e.g., time-gating and null distance) and advanced photonic components (e.g., vertical cavity source-emitting laser as light sources, single photon avalanche diode, and silicon photomultipliers as detectors, fast-gating circuits, and time-to-digital converters for acquisition) are focused. This study shows how these tools could lead on one hand to compact and wearable time-domain devices for point-of-care diagnostics down to the consumer level and on the other hand to powerful systems with exceptional depth penetration and sensitivity.

Diffuse optical imaging and spectroscopy of the female breast is an area of active research. We review the present status of this field and discuss the broad range of methodologies and applications. Starting with a brief overview on breast physiology, the remodeling of vasculature and extracellular matrix caused by solid tumors is highlighted that is relevant for contrast in optical imaging. Then, the various instrumental techniques and the related methods of data analysis and image generation are described and compared including multimodality instrumentation, fluorescence mammography, broadband spectroscopy, and diffuse correlation spectroscopy. We review the clinical results on functional properties of malignant and benign breast lesions compared to host tissue and discuss the various methods to improve contrast between healthy and diseased tissue, such as enhanced spectroscopic information, dynamic variations of functional properties, pharmacokinetics of extrinsic contrast agents, including the enhanced permeability and retention effect. We discuss research on monitoring neoadjuvant chemotherapy and on breast cancer risk assessment as potential clinical applications of optical breast imaging and spectroscopy. Moreover, we consider new experimental approaches, such as photoacoustic imaging and long-wavelength tissue spectroscopy.

Near-infrared diffuse optical tomography (DOT), one of the most sophisticated optical imaging techniques for observations through biological tissue, allows 3-D quantitative imaging of optical properties, which include functional and anatomical information. With DOT, it is expected to be possible to overcome the limitations of conventional near-infrared spectroscopy (NIRS) as well as offering the potential for diagnostic optical imaging. However, DOT has been under development for more than 30 years, and the difficulties in development are attributed to the fact that light is strongly scattered and that diffusive photons are used for the image reconstruction. The DOT algorithm is based on the techniques of inverse problems. The radiative transfer equation accurately describes photon propagation in biological tissue, while, because of its high computation load, the diffusion equation (DE) is often used as the forward model. However, the DE is invalid in low-scattering and/or highly absorbing regions and in the vicinity of light sources. The inverse problem is inherently ill-posed and highly undetermined. Here, we first summarize NIRS and then describe various approaches in the efforts to develop accurate and efficient DOT algorithms and present some examples of clinical applications. Finally, we discuss the future prospects of DOT.

In most daily activities related to work or leisure, the energy for muscle work substantially comes from oxidative metabolism. Functional limitations or impairments of this metabolism can significantly affect exercise tolerance and performance. As a method for the functional evaluation of skeletal muscle oxidative metabolism, near-infrared spectroscopy (NIRS) has important strengths but also several limitations, some of which have been overcome by recent technological developments. Skeletal muscle fractional O₂ extraction, the main variable which can be noninvasively evaluated by NIRS, is the result of the dynamic balance between O₂ utilization and O₂ delivery; it can yield relevant information on key physiological and pathophysiological mechanisms, relevant in the evaluation of exercise performance and exercise tolerance in healthy subjects (in normal and in altered environmental conditions) and in patients. In the right hands, NIRS can offer insights into the physiological and pathophysiological adaptations to conditions of increased O₂ needs that involve, in an integrated manner, different organs and systems of the body. In terms of patient evaluation, NIRS allows determination of the evolution of the functional impairments, to identify their correlations with clinical symptoms, to evaluate the effects of therapeutic or rehabilitative interventions, and to gain pathophysiological and diagnostic insights.

Ambulatory diffuse optical tomography (aDOT) is based on near-infrared spectroscopy (NIRS) and enables three-dimensional imaging of regional hemodynamics and oxygen consumption during a person’s normal activities. Although NIRS has been previously used for muscle assessment, it has been notably limited in terms of the number of channels measured, the extent to which subjects can be ambulatory, and/or the ability to simultaneously acquire synchronized auxiliary data such as electromyography (EMG) or electrocardiography (ECG). We describe the development of a prototype aDOT system, called NINscan-M, capable of ambulatory tomographic imaging as well as simultaneous auxiliary multimodal physiological monitoring. Powered by four AA size batteries and weighing 577 g, the NINscan-M prototype can synchronously record 64-channel NIRS imaging data, eight channels of EMG, ECG, or other analog signals, plus force, acceleration, rotation, and temperature for 24+ h at up to 250 Hz. We describe the system’s design, characterization, and performance characteristics. We also describe examples of isometric, cycle ergometer, and free-running ambulatory exercise to demonstrate tomographic imaging at 25 Hz. NINscan-M represents a multiuse tool for muscle physiology studies as well as clinical muscle assessment.

Brain functional activity involves complex cellular, metabolic, and vascular chain reactions, making it difficult to comprehend. Electroencephalography (EEG) and functional near infrared spectroscopy (fNIRS) have been combined into a multimodal neuroimaging method that captures both electrophysiological and hemodynamic information to explore the spatiotemporal characteristics of brain activity. Because of the significance of visually evoked functional activity in clinical applications, numerous studies have explored the amplitude of the visual evoked potential (VEP) to clarify its relationship with the hemodynamic response. However, relatively few studies have investigated the influence of latency, which has been frequently used to diagnose visual diseases, on the hemodynamic response. Moreover, because the latency and the amplitude of VEPs have different roles in coding visual information, investigating the relationship between latency and the hemodynamic response should be helpful. In this study, checkerboard reversal tasks with graded contrasts were used to evoke visual functional activity. Both EEG and fNIRS were employed to investigate the relationship between neuronal electrophysiological activities and the hemodynamic responses. The VEP amplitudes were linearly correlated with the hemodynamic response, but the VEP latency showed a negative linear correlation with the hemodynamic response.

Optically derived tissue properties across a range of breast densities and the effects of breast compression on estimates of hemoglobin, oxygen metabolism, and water and lipid concentrations were obtained from a coregistered imaging system that integrates near-infrared spectral tomography (NIRST) with digital breast tomosynthesis (DBT). Image data were analyzed from 27 women who underwent four IRB approved NIRST/DBT exams that included fully and mildly compressed breast acquisitions in two projections—craniocaudal (CC) and mediolateral-oblique (MLO)—and generated four data sets per patient (full and moderate compression in CC and MLO views). Breast density was correlated with HbT (r=0.64, p=0.001), water (r=0.62, p=0.003), and lipid concentrations (r=−0.74, p<0.001), but not oxygen saturation. CC and MLO views were correlated for individual subjects and demonstrated no statistically significant differences in grouped analysis. Comparison of compressed and uncompressed imaging demonstrated a significant decrease in oxygen saturation under compression (58% versus 50%, p=0.04). Mammographic breast density categorization was correlated with measured optically derived properties.

Laser speckle imaging (LSI) enables measurement of relative changes in blood flow in biological tissues. We postulate that a point-of-care form factor will lower barriers to routine clinical use of LSI. Here, we describe a first-generation handheld LSI device based on a tablet computer. The coefficient of variation of speckle contrast was <2% after averaging imaging data collected over an acquisition period of 5.3 s. With a single, experienced user, handheld motion artifacts had a negligible effect on data collection. With operation by multiple users, we did not identify any significant difference (p<0.05) between the measured speckle contrast values using either a handheld or mounted configuration. In vivo data collected during occlusion experiments demonstrate that a handheld LSI is capable of both quantitative and qualitative assessment of changes in blood flow. Finally, as a practical application of handheld LSI, we collected data from a 53-day-old neonate with confirmed compromised blood flow in the hand. We readily identified with LSI a region of diminished blood flow in the thumb of the affected hand. Our data collectively suggest that handheld LSI is a promising technique to enable clinicians to obtain point-of-care measurements of blood flow.

Invasive brain cancer cells cannot be visualized during surgery and so they are often not removed. These residual cancer cells give rise to recurrences. In vivo Raman spectroscopy can detect these invasive cancer cells in patients with grade 2 to 4 gliomas. The robustness of this Raman signal can be dampened by spectral artifacts generated by lights in the operating room. We found that artificial neural networks (ANNs) can overcome these spectral artifacts using nonparametric and adaptive models to detect complex nonlinear spectral characteristics. Coupling ANN with Raman spectroscopy simplifies the intraoperative use of Raman spectroscopy by limiting changes required to the standard neurosurgical workflow. The ability to detect invasive brain cancer under these conditions may reduce residual cancer remaining after surgery and improve patient survival.

Chromosome ends are shielded from exonucleolytic attack and inappropriate end-joining by terminal structures called telomeres; these structures are potential targets for anticancer drugs. Telomeres are composed of a simple DNA sequence (5′-TTAGGG-3′ in humans) repeated more than a thousand times, a short 3′ single-stranded overhang, and numerous proteins. Electron microscopy has shown that the 3′ overhang pairs with the complementary strand at an internal site creating a small displacement loop and a large double-stranded “t-loop.” Our goal is to determine whether all telomeres adopt the t-loop configuration, or whether there are two or more distinct configurations. Progress in optimizing super-resolution (SR) microscopy for this ongoing investigation is reported here. Results suggest that under certain conditions sample preparation procedures may disrupt chromatin by causing loss of nucleosomes. This finding may limit the use of SR microscopy in telomere studies.

We performed ratiometric analysis of retinal optical coherence tomography images for the first time in multiple sclerosis (MS) patients. The ratiometric analysis identified differences in several retinal layer thickness ratios in the cohort of MS subjects without a history of optic neuritis (ON) compared to healthy control (HC) subjects, and there was no difference in standard retinal nerve fiber layer thickness (RNFL_T). The difference in such ratios between HC subjects and those with mild MS-disability, without a difference in RNFL_T, further suggests the possibility of using layer ratiometric analysis for detecting early retinal changes in MS. Ratiometric analysis may be useful and potentially more sensitive for detecting disease changes in MS.

Utilizing a laser scanning confocal microscope system, the refractive indices of articular cartilage (AC) with mechanical or biochemical degenerations were characterized to investigate whether potential correlations exist between refractive index (RI) and cartilage degeneration. The cartilage samples collected from the medial femoral condyles of kangaroo knees were mechanically degenerated under different loading patterns or digested in trypsin solution with different concentrations. The sequences of RI were then measured from cartilage surface to deep region and the fluctuations of RI were quantified considering combined effects of fluctuating frequency and amplitude. The compositional and microstructural alterations of cartilage samples were assessed with histological methods. Along with the loss of proteoglycans, the average RI of cartilage increased and the local fluctuation of RI became stronger. Short-term high-speed test induced little influence to both the depth fluctuation and overall level of RI. Long-term low-speed test increased the fluctuation of RI but the average RI was barely changed. The results substantially demonstrate that RI of AC varies with both compositional and structural alterations and is potentially an indicator for the degeneration of AC.

We propose and objectively evaluate an inverse Monte Carlo model for estimation of absorption and reduced scattering coefficients and similarity parameter γ from spatially resolved reflectance (SRR) profiles in the subdiffusive regime. The similarity parameter γ carries additional information on the phase function that governs the angular properties of scattering in turbid media. The SRR profiles at five source-detector separations were acquired with an optical fiber probe. The inverse Monte Carlo model was based on a cost function that enabled robust estimation of optical properties from a few SRR measurements without a priori knowledge about spectral dependencies of the optical properties. Validation of the inverse Monte Carlo model was performed on synthetic datasets and measured SRR profiles of turbid phantoms comprising molecular dye and polystyrene microspheres. We observed that the additional similarity parameter γ substantially reduced the reflectance variability arising from the phase function properties and significantly improved the accuracy of the inverse Monte Carlo model. However, the observed improvement of the extended inverse Monte Carlo model was limited to reduced scattering coefficients exceeding ∼15  cm⁻¹, where the relative root-mean-square errors of the estimated optical properties were well within 10%.

Spatial variation of refractive index (RI) in connective tissues exhibits multifractality, which encodes useful morphological and ultrastructural information about the disease. We present a spectral Mueller matrix (MM)-based approach in combination with multifractal detrended fluctuation analysis (MFDFA) to exclusively pick out the signature of the underlying connective tissue multifractality through the superficial epithelium layer. The method is based on inverse analysis on selected spectral scattering MM elements encoding the birefringence information on the anisotropic connective tissue. The light scattering spectra corresponding to the birefringence carrying MM elements are then subjected to the Born approximation-based Fourier domain preprocessing to extract ultrastructural RI fluctuations of anisotropic tissue. The extracted RI fluctuations are subsequently analyzed via MFDFA to yield the multifractal tissue parameters. The approach was experimentally validated on a simple tissue model comprising of TiO₂ as scatterers of the superficial isotropic layer and rat tail collagen as an underlying anisotropic layer. Finally, the method enabled probing of precancer-related subtle alterations in underlying connective tissue ultrastructural multifractality from intact tissues.

This study demonstrates the evaluation of Raman spectroscopy as a rapid diagnostic test in comparison to commonly performed tests for an accurate detection of dengue fever in human blood sera. Blood samples of 104 suspected dengue patients collected from Holy Family Hospital, Rawalpindi, Pakistan, have been used in this study. Out of 104 samples, 52 (50%) were positive based on immunoglobulin G (IgG), whereas 54 (52%) were positive based on immunoglobulin M (IgM) antibody tests. For the determination of the diagnostic capabilities of Raman spectroscopy, accuracy, sensitivity, specificity and false positive rate have been calculated in comparison to normally performed IgM and IgG captured enzyme-linked immunosorbent assay tests. Accuracy, precision, specificity, and sensitivity for Raman spectroscopy in comparison to IgM were found to be 66%, 70%, 72%, and 61%, whereas based on IgG they were 47%, 46%, 52%, and 43%, respectively.

We describe a systematic approach to image, track, and quantify the movements of HIV viruses embedded in human cervical mucus. The underlying motivation for this study is that, in HIV-infected adults, women account for more than half of all new cases and most of these women acquire the infection through heterosexual contact. The endocervix is believed to be a susceptible site for HIV entry. Cervical mucus, which coats the endocervix, should play a protective role against the viruses. Thus, we developed a methodology to apply time-resolved confocal microscopy to examine the motion of HIV viruses that were added to samples of untreated cervical mucus. From the images, we identified the viruses, tracked them over time, and calculated changes of the statistical mean-squared displacement (MSD) of each virus. Approximately half of tracked viruses appear constrained while the others show mobility with MSDs that are proportional to τ^α+ν²τ², over time range τ, depicting a combination of anomalous diffusion (0<α<0.40) and flow-like behavior. The MSD data also reveal plateaus attributable to possible stalling of the viruses. Although a more extensive study is warranted, these results support the assumption of mucus being a barrier against the motion of these viruses.

The great interest in upconversion nanoparticles exists due to their high efficiency under multiphoton excitation. However, when these particles are used in scanning microscopy, the upconversion luminescence causes a streaking effect due to the long lifetime. This article describes a method of upconversion microparticle luminescence lifetime determination with help of modified Lucy–Richardson deconvolution of laser scanning microscope (LSM) image obtained under near-IR excitation using nondescanned detectors. Determination of the upconversion luminescence intensity and the decay time of separate microparticles was done by intensity profile along the image fast scan axis approximation. We studied upconversion submicroparticles based on fluoride hosts doped with Yb³⁺-Er³⁺ and Yb³⁺-Tm³⁺ rare earth ion pairs, and the characteristic decay times were 0.1 to 1.5 ms. We also compared the results of LSM measurements with the photon counting method results; the spread of values was about 13% and was associated with the approximation error. Data obtained from live cells showed the possibility of distinguishing the position of upconversion submicroparticles inside and outside the cells by the difference of their lifetime. The proposed technique allows using the upconversion microparticles without shells as probes for the presence of OH− ions and CO₂ molecules.

Early detection of the most prevalent oral disease worldwide, i.e., dental caries, still remains as one of the major challenges in dentistry. The current dental standard of care relies on caries detection methods, such as visual inspection and x-ray radiography, which lack the sufficient specificity and sensitivity to detect caries at early stages of formation when they can be healed. We report on the feasibility of early caries detection in a clinically and commercially viable thermophotonic imaging system. The system incorporates intensity-modulated laser light along with a low-cost long-wavelength infrared (LWIR; 8 to 14  μm) camera, providing diagnostic contrast based on the enhanced light absorption of early caries. The LWIR camera is highly suitable for integration into clinical platforms because of its low weight and cost. In addition, through theoretical modeling, we show that LWIR detection enhances the diagnostic contrast due to the minimal LWIR transmittance of enamel and suppression of the masking effect of the direct thermal Planck emission. Diagnostic performance of the system and its detection threshold are experimentally evaluated by monitoring the inception and progression of artificially induced occlusal and smooth surface caries. The results are suggestive of the suitability of the developed LWIR system for detecting early dental caries.

Optical coherence tomography (OCT) enables us to monitor alterations in the thickness of the retinal layer as disease progresses in the human retina. However, subtle morphological changes in the retinal layers due to early disease progression often may not lead to detectable alterations in the thickness. OCT images encode depth-dependent backscattered intensity distribution arising due to the depth distributions of the refractive index from tissue microstructures. Here, such depth-resolved refractive index variations of different retinal layers were analyzed using multifractal detrended fluctuation analysis, a special class of multiresolution analysis tools. The analysis extracted and quantified microstructural multifractal information encoded in normal as well as diseased human retinal OCT images acquired in vivo. Interestingly, different layers of the retina exhibited different degrees of multifractality in a particular retina, and the individual layers displayed consistent multifractal trends in healthy retinas of different human subjects. In the retinal layers of diabetic macular edema (DME) subjects, the change in multifractality manifested prominently near the boundary of the DME as compared to the normal retinal layers. The demonstrated ability to quantify depth-resolved information on multifractality encoded in OCT images appears promising for the early diagnosis of diseases of the human eye, which may also prove useful for detecting other types of tissue abnormalities from OCT images.

Scattering within biological samples limits the imaging depth and the resolution in microscopy. We present a prior and regularization approach for blind deconvolution algorithms to correct the influence of scattering to increase the imaging depth and resolution. The effect of the prior is demonstrated on a three-dimensional image stack of a zebrafish embryo captured with a selective plane illumination microscope. Blind deconvolution algorithms model the recorded image as a convolution between the distribution of fluorophores and a point spread function (PSF). Our prior uses image information from adjacent z-planes to estimate the unknown blur in tissue. The increased size of the PSF due to the cascading effect of scattering in deeper tissue is accounted for by a depth adaptive regularizer model. In a zebrafish sample, we were able to extend the point in depth, where scattering has a significant effect on the image quality by around 30  μm.

Adequate assessment of burn wounds is crucial in the management of burn patients. Thermography, as a noninvasive measurement tool, can be utilized to detect the remaining perfusion over large burn wound areas by measuring temperature, thereby reflecting the healing potential (HP) (i.e., number of days that burns require to heal). The objective of this study was to evaluate the clinimetric properties (i.e., reliability and validity) of thermography for measuring burn wound HP. To evaluate reliability, two independent observers performed a thermography measurement of 50 burns. The intraclass correlation coefficient (ICC), the standard error of measurement (SEM), and the limits of agreement (LoA) were calculated. To assess validity, temperature differences between burned and nonburned skin (ΔT) were compared to the HP found by laser Doppler imaging (serving as the reference standard). By applying a visual method, one ΔT cutoff point was identified to differentiate between burns requiring conservative versus surgical treatment. The ICC was 0.99, expressing an excellent correlation between two measurements. The SEM was calculated at 0.22°C, the LoA at −0.58°C and 0.64°C. The ΔT cutoff point was −0.07°C (sensitivity 80%; specificity 80%). These results show that thermography is a reliable and valid technique in the assessment of burn wound HP.

Retinal image quality assessment (RIQA) is an essential step in automated screening systems to avoid misdiagnosis caused by processing poor quality retinal images. A no-reference transform-based RIQA algorithm is introduced that assesses images based on five clarity and content quality issues: sharpness, illumination, homogeneity, field definition, and content. Transform-based RIQA algorithms have the advantage of considering retinal structures while being computationally inexpensive. Wavelet-based features are proposed to evaluate the sharpness and overall illumination of the images. A retinal saturation channel is designed and used along with wavelet-based features for homogeneity assessment. The presented sharpness and illumination features are utilized to assure adequate field definition, whereas color information is used to exclude nonretinal images. Several publicly available datasets of varying quality grades are utilized to evaluate the feature sets resulting in area under the receiver operating characteristic curve above 0.99 for each of the individual feature sets. The overall quality is assessed by a classifier that uses the collective features as an input vector. The classification results show superior performance of the algorithm in comparison to other methods from literature. Moreover, the algorithm addresses efficiently and comprehensively various quality issues and is suitable for automatic screening systems.

Tissues are characterized by a strong scattering of visible optical radiation, which prevents one from achieving deep-tissue imaging. We propose a computational imaging technique for the inference of specific macroscopic, spatial phase distribution features of the scattering media. The spatial phase distribution is reconstructed from several defocused intensity images. We empirically demonstrate the method by reconstructing the location of two fibula chicken bones, embedded within chicken breast tissue. The suggested technique is safe, using visible laser illumination, and noninvasive. It is also cost-effective since a simple optical system is used and the images are acquired using a conventional camera, and it does not require interferometric detection as well as direct access to the object in absence of the layer.

Structured illumination microscopy (SIM) is an attractive choice for fast superresolution imaging. The generation of structured illumination patterns made by interference of laser beams is broadly employed to obtain high modulation depth of patterns, while the polarizations of the laser beams must be elaborately controlled to guarantee the high contrast of interference intensity, which brings a more complex configuration for the polarization control. The emerging pattern projection strategy is much more compact, but the modulation depth of patterns is deteriorated by the optical transfer function of the optical system, especially in high spatial frequency near the diffraction limit. Therefore, the traditional superresolution reconstruction algorithm for interference-based SIM will suffer from many artifacts in the case of projection-based SIM that possesses a low modulation depth. Here, we propose an alternative reconstruction algorithm based on image recombination transform, which provides an alternative solution to address this problem even in a weak modulation depth. We demonstrated the effectiveness of this algorithm in the multicolor superresolution imaging of bovine pulmonary arterial endothelial cells in our developed projection-based SIM system, which applies a computer controlled digital micromirror device for fast fringe generation and multicolor light-emitting diodes for illumination. The merit of the system incorporated with the proposed algorithm allows for a low excitation intensity fluorescence imaging even less than 1  W/cm², which is beneficial for the long-term, in vivo superresolved imaging of live cells and tissues.

Intrinsic optical signal (IOS) imaging promises a noninvasive method for advanced study and diagnosis of eye diseases. Before pursuing clinical applications, it is essential to understand anatomic and physiological sources of retinal IOSs and to establish the relationship between IOS distortions and eye diseases. The purpose of this study was designed to demonstrate the feasibility of in vivo IOS imaging of mouse models. A high spatiotemporal resolution spectral domain optical coherence tomography (SD-OCT) was employed for depth-resolved retinal imaging. A custom-designed animal holder equipped with ear bar and bite bar was used to minimize eye movements. Dynamic OCT imaging revealed rapid IOS from the photoreceptor’s outer segment immediately after the stimulation delivery, and slow IOS changes were observed from inner retinal layers. Comparative photoreceptor IOS and electroretinography recordings suggested that the fast photoreceptor IOS may be attributed to the early stage of phototransduction before the hyperpolarization of retinal photoreceptor.

Studies have proven the relationship between cutaneous vasculature abnormalities and dermatological disorders, but to image vasculature noninvasively in vivo, advanced optical imaging techniques are required. In this study, we imaged a palm of a healthy volunteer and three subjects with cutaneous abnormalities with photoacoustic tomography (PAT) and optical coherence tomography with angiography extension (OCTA). Capillaries in the papillary dermis that are too small to be discerned with PAT are visualized with OCTA. From our results, we speculate that the PA signal from the palm is mostly from hemoglobin in capillaries rather than melanin, knowing that melanin concentration in volar skin is significantly smaller than that in other areas of the skin. We present for the first time OCTA images of capillaries along with the PAT images of the deeper vessels, demonstrating the complementary effective imaging depth range and the visualization capabilities of PAT and OCTA for imaging human skin in vivo. The proposed imaging system in this study could significantly improve treatment monitoring of dermatological diseases associated with cutaneous vasculature abnormalities.

We report a fluorescent probe for mRNA detection. It consists of a gold nanorod (GNR) functionalized with fluorophore-labeled hairpin oligonucleotides (hpDNA) that are complementary to the mRNA of a target gene. This nanoprobe was found to be sensitive to a complementary oligonucleotide, as indicated by significant changes in both fluorescence intensity and lifetime. The influence of the surface density of hpDNA on the performance of this nanoprobe was investigated, suggesting that high hybridization efficiency could be achieved at a relatively low surface loading density of hpDNA. However, steady-state fluorescence spectroscopy revealed better overall performance, in terms of sensitivity and detection range, for nanoprobes with higher hairpin coverage. Time-resolved fluorescence lifetime spectroscopy revealed significant lifetime changes of the fluorophore upon hybridization of hpDNA with targets, providing further insight on the hybridization kinetics of the probe as well as the quenching efficiency of GNRs.

An adequate water balance (hydration extent) is one of the basic factors of normal eye function, including its external shells: the cornea and the sclera. Adequate control of corneal and scleral hydration is very important for early diagnosis of a variety of eye diseases, stating indications for and contraindications against keratorefractive surgeries and the choice of contact lens correction solutions. THz systems of creating images in reflected beams are likely to become ideal instruments of noninvasive control of corneal and scleral hydration degrees. This paper reports on the results of a study involving transmittance and reflectance spectra for the cornea and the sclera of rabbit and human eyes, as well as those of the rabbit eye, in the frequency range of 0.13 to 0.32 THz. The dependence of the reflectance coefficient of these tissues on water mass percentage content was determined. The experiments were performed on three corneas, three rabbit scleras, two rabbit eyes, and three human scleras. The preliminary results demonstrate that the proposed technique, based on the use of a continuous THz radiation, may be utilized to create a device for noninvasive control of corneal and scleral hydration, which has clear potential of broad practical application.

This study aimed to compare two tissue oximeters, the INVOS 5100c and the Equanox 7600, in terms of their reproducibility and the interchangeability of their measures. In a randomized order, three measurements were taken at six different sites on both sides of the body in 53 healthy volunteers. Intraclass correlation coefficients (ICC) and within-subject standard deviation (S_w) were calculated for each device. The ICCs were compared using Fisher r-to-z transformation and the S_w were compared using paired-sample t-tests. We found no difference between the reproducibility of the INVOS {ICC=0.92 [95% confidence interval (CI) 0.90 to 0.93]} and Equanox [ICC=0.90 (95% CI 0.88 to 0.93)] in terms of ICCs (p=0.06). However, the Equanox [S_w=1.96 (95% CI 1.91 to 2.02)] showed a better S_w than the INVOS [S_w=2.11 (95% CI 2.05 to 2.17)] (p=0.019). Also, when compared directly to stable condition, the readings produced by the two oximeters varied considerably [ICC 0.43 (95% CI 0.36 to 0.49)]. When taken individually, both tissue oximeters displayed good reproducibility, the Equanox being slightly better than the INVOS in terms of absolute reproducibility. However, when compared, the oximeters showed poor interdevices agreement. Reference values were also described.

There is a strong need to develop clinical instruments that can perform rapid tissue assessment at the tip of smart clinical instruments for a variety of oncological applications. This study presents the first in vivo real-time tissue characterization during 24 liver biopsy procedures using diffuse reflectance (DR) spectroscopy at the tip of a core biopsy needle with integrated optical fibers. DR measurements were performed along each needle path, followed by biopsy of the target lesion using the same needle. Interventional imaging was coregistered with the DR spectra. Pathology results were compared with the DR spectroscopy data at the final measurement position. Bile was the primary discriminator between normal liver tissue and tumor tissue. Relative differences in bile content matched with the tissue diagnosis based on histopathological analysis in all 24 clinical cases. Continuous DR measurements during needle insertion in three patients showed that the method can also be applied for biopsy guidance or tumor recognition during surgery. This study provides an important validation step for DR spectroscopy-based tissue characterization in the liver. Given the feasibility of the outlined approach, it is also conceivable to make integrated fiber-optic tools for other clinical procedures that rely on accurate instrument positioning.

Cupriavidus necator accumulates large amounts of poly(3-hydroxybutyrate) (PHB), a biodegradable substitute for petroleum-based plastics, under certain nutrient conditions. Conventional solvent-extraction-based methods for PHB quantification only obtain average information from cell populations and, thus, mask the heterogeneity among individual cells. Laser tweezers Raman spectroscopy (LTRS) was used to monitor dynamic changes in the contents of PHB, nucleic acids, and proteins in C. necator at the population and single-cell levels when the microorganism cells were cultivated at various carbon-to-nitrogen ratios. The biosynthetic activities of nucleic acids and proteins were maintained at high levels, and only a small amount of PHB was produced when the bacterial cells were cultured under balanced growth conditions. By contrast, the syntheses of nucleic acids and proteins were blocked, and PHB was accumulated in massive amount inside the microbial cells under nitrogen-limiting growth circumstances. Single-cell analysis revealed a relatively high heterogeneity in PHB level at the early stage of the bacterial growth. Additionally, bacterial cells in populations at certain cultivation stages were composed of two or three subpopulations on the basis of their PHB abundance. Overall, LTRS is a reliable single-cell analysis tool that can provide insights into PHB fermentation.

Laser photocoagulation is an established treatment for a variety of retinal diseases. However, when using the same irradiation parameter, the size and strength of the lesions are unpredictable due to unknown inter- and intraindividual optical properties of the fundus layers. The aim of this work is to investigate a feedback system to generate desired lesions of preselectable strengths by automatically controlling the irradiation time. Optoacoustics were used for retinal temperature monitoring. A 532-nm continuous wave Nd:YAG laser was used for photocoagulation. A 75-ns/523-nm Q-switched Nd:YLF laser simultaneously excited temperature-dependent pressure transients, which were detected at the cornea by an ultrasonic transducer embedded in a contact lens. The temperature data were analyzed during the irradiation by a LabVIEW routine. The treatment laser was switched off automatically when the required lesion strength was achieved. Five different feedback control algorithms for different lesion sizes were developed and tested on rabbits in vivo. With a laser spot diameter of 133  μm, five different lesion types with ophthalmoscopically visible diameters ranging mostly between 100 and 200  μm, and different appearances were achieved by automatic exposure time control. The automatically controlled lesions were widely independent of the treatment laser power and the retinal pigmentation.

The purpose of this study was to evaluate the influences of helium–neon (He–Ne) and infrared (IR) lasers on the viability and proliferation rate of healthy and ovariectomy-induced osteoporotic (OVX) bone marrow mesenchymal stem cells (BMMSCs) in vitro. MSCs harvested from the BM of healthy and OVX rats were culture expanded. He–Ne and IR lasers were applied three times at energy densities of 0.6, 1.2, and 2.4  J/cm² for BMMSCs. BMMSCs viability and proliferation rate were evaluated by MTT assay on days 2, 4, 6, 14, and 21. The results showed that healthy BMMSCs responded optimally to 0.6  J/cm² using an IR laser after three times of laser radiation. Moreover, it was found that OVX-BMMSCs responded optimally to 0.6  J/cm² with He–Ne laser and one-time laser radiation. It is concluded that the low-level laser therapy (LLLT) effect depends on the physiological state of the BMMSCs, type of the laser, wavelength, and number of laser sessions. The biostimulation efficiency of LLLT also depends on the delivered energy density. LLLT can enhance the viability and proliferation rate of healthy and especially osteoporotic autologous BMMSCs, which could be very useful in regenerative medicine.

This PDF file contains the errata for “JBO Vol. 21 Issue 09 Paper JBO-2016-0524-ERR” for JBO Vol. 21 Issue 09