Two-photon fluorescence (TPF) and second harmonic generation (SHG) microscopy provide direct visualization of the skin dermal fibers in vivo. A typical method for analyzing TPF/SHG images involves averaging the image intensity and therefore disregarding the spatial distribution information. The goal of this study is to develop an algorithm to document age-related effects of the dermal matrix. TPF and SHG images were acquired from the upper inner arm, volar forearm, and cheek of female volunteers of two age groups: 20 to 30 and 60 to 80 years of age. The acquired images were analyzed for parameters relating to collagen and elastin fiber features, such as orientation and density. Both collagen and elastin fibers showed higher anisotropy in fiber orientation for the older group. The greatest difference in elastin fiber anisotropy between the two groups was found for the upper inner arm site. Elastin fiber density increased with age, whereas collagen fiber density decreased with age. The proposed analysis considers the spatial information inherent to the TPF and SHG images and provides additional insights into how the dermal fiber structure is affected by the aging process.

Saturation artifacts that are commonly observed in endoscopic swept-source optical coherence tomography (SSOCT) images cause image degradation and loss of image information. We present work on the correction of saturation effects in endoscopic SSOCT imaging. This method utilizes a broadband power divider with excellent pick-off flatness to divide the detected interference signal into the two channels of an analog-to-digital converter. Based on the precise calibration of the splitting ratio between the two channels, the maximum measurable signal power of the system was drastically increased by using the low level signal in one channel to correct the saturated signal in the other channel. The experimental results demonstrated that this technique can efficiently correct the saturation artifacts in endoscopic two- and three-dimensional SSOCT images in an accurate and cost-effective manner.

We quantitatively investigated the measurement sensitivity of spatially resolved spectroscopy (SRS) across six tissue models: cerebral tissue, a small animal brain, the forehead of a fetus, an adult brain, forearm muscle, and thigh muscle. The optical path length in the voxel of the model was analyzed using Monte Carlo simulations. It was found that the measurement sensitivity can be represented as the product of the change in the absorption coefficient and the difference in optical path length in two states with different source–detector distances. The results clarified the sensitivity ratio between the surface layer and the deep layer at each source–detector distance for each model and identified changes in the deep measurement area when one of the detectors was close to the light source. A comparison was made with the results from continuous-wave spectroscopy. The study also identified measurement challenges that arise when the surface layer is inhomogeneous. Findings on the measurement sensitivity of SRS at each voxel and in each layer can support the correct interpretation of measured values when near-infrared oximetry or functional near-infrared spectroscopy is used to investigate different tissue structures.

Radiation therapy produces Cherenkov optical emission in tissue, and this light can be utilized to activate molecular probes. The feasibility of sensing luminescence from a tissue molecular oxygen sensor from within a human body phantom was examined using the geometry of the axillary lymph node region. Detection of regions down to 30-mm deep was feasible with submillimeter spatial resolution with the total quantity of the phosphorescent sensor PtG4 near 1 nanomole. Radiation sheet scanning in an epi-illumination geometry provided optimal coverage, and maximum intensity projection images provided illustration of the concept. This work provides the preliminary information needed to attempt this type of imaging in vivo.

Rare-earth-doped nanocomposites have appealing optical properties for use as biomedical contrast agents, but few systems exist for imaging these materials. We describe the design and characterization of (i) a preclinical system for whole animal in vivo imaging and (ii) an integrated optical coherence tomography/confocal microscopy system for high-resolution imaging of ex vivo tissues. We demonstrate these systems by administering erbium-doped nanocomposites to a murine model of metastatic breast cancer. Short-wave infrared emissions were detected in vivo and in whole organ imaging ex vivo. Visible upconversion emissions and tissue autofluorescence were imaged in biopsy specimens, alongside optical coherence tomography imaging of tissue microstructure. We anticipate that this work will provide guidance for researchers seeking to image these nanomaterials across a wide range of biological models.

We report a photoacoustic computed tomography (PACT) system using a customized optical fiber with a cylindrical diffuser to internally illuminate deep targets. The traditional external light illumination in PACT usually limits the penetration depth to a few centimeters from the tissue surface, mainly due to strong optical attenuation along the light propagation path from the outside in. By contrast, internal light illumination, with external ultrasound detection, can potentially detect much deeper targets. Different from previous internal illumination PACT implementations using forward-looking optical fibers, our internal-illumination PACT system uses a customized optical fiber with a 3-cm-long conoid needle diffuser attached to the fiber tip, which can homogeneously illuminate the surrounding space and substantially enlarge the field of view. We characterized the internal illumination distribution and PACT system performance. We performed tissue phantom and in vivo animal studies to further demonstrate the superior imaging depth using internal illumination over external illumination. We imaged a 7.5-cm-deep leaf target embedded in optically scattering medium and the beating heart of a mouse overlaid with 3.7-cm-thick chicken tissue. Our results have collectively demonstrated that the internal light illumination combined with external ultrasound detection might be a useful strategy to improve the penetration depth of PACT in imaging deep organs of large animals and humans.

Medical devices face many hurdles before they enter routine clinical practice to address unmet clinical needs. This is also the case for biomedical optical spectroscopy and imaging systems that are used here to illustrate the opportunities and challenges involved. Following initial concept, stages in clinical translation include instrument development, preclinical testing, clinical prototyping, clinical trials, prototype-to-product conversion, regulatory approval, commercialization, and finally clinical adoption and dissemination, all in the face of potentially competing technologies. Optical technologies face additional challenges from their being extremely diverse, often targeting entirely different diseases and having orders-of-magnitude differences in resolution and tissue penetration. However, these technologies can potentially address a wide variety of unmet clinical needs since they provide rich intrinsic biochemical and structural information, have high sensitivity and specificity for disease detection and localization, and are practical, safe (minimally invasive, nonionizing), and relatively affordable.

A set of methods in terms of both image processing and gaze estimation for accurate eye tracking is proposed. The eye-tracker used in this study relies on the dark-pupil method with up to 12 corneal reflections and offers an unprecedented high resolution imaging of the pupil and the cornea. The potential benefits of a higher number of glints and their optimum arrangement are analyzed considering distinct light sources configurations with 12, 8, 6, 4, and 2 corneal reflections. Moreover, a normalization factor of the pupil-glint vector is proposed for each configuration. There is a tendency for increasing accuracy with the number of glints, especially vertically (0.47 deg for 12 glints configuration versus 0.65 deg for 2 glints configuration). Besides the number of corneal reflections, their arrangement seems to have a stronger effect. A configuration that minimizes the interference of the eyelids with the corneal reflections is desired. Finally, the normalization of the pupil-glint vectors improves the vertical eye tracking accuracy up to 43.2%. In addition, the normalization also limits the need for a higher number of light sources to achieve better spatial accuracy.

Caspase-3 is known for its role in apoptosis and programmed cell death regulation. We detected caspase-3 activation in vivo in tumor xenografts via shift of mean fluorescence lifetimes of a caspase-3 sensor. We used the genetically encoded sensor TR23K based on the red fluorescent protein TagRFP and chromoprotein KFP linked by 23 amino acid residues (TagRFP-23-KFP) containing a specific caspase cleavage DEVD motif to monitor the activity of caspase-3 in tumor xenografts by means of fluorescence lifetime imaging-Forster resonance energy transfer. Apoptosis was induced by injection of paclitaxel for A549 lung adenocarcinoma and etoposide and cisplatin for HEp-2 pharynx adenocarcinoma. We observed a shift in lifetime distribution from 1.6 to 1.9 ns to 2.1 to 2.4 ns, which indicated the activation of caspase-3. Even within the same tumor, the lifetime varied presumably due to the tumor heterogeneity and the different depth of tumor invasion. Thus, processing time-resolved fluorescence images allows detection of both the cleaved and noncleaved states of the TR23K sensor in real-time mode during the course of several weeks noninvasively. This approach can be used in drug screening, facilitating the development of new anticancer agents as well as improvement of chemotherapy efficiency and its adaptation for personal treatment.

Intraocular lenses (IOLs) are used in the cataract treatment for surgical replacement of the opacified crystalline lens. Before being implanted they have to pass the strict quality control to guarantee a good biomechanical stability inside the capsular bag, avoiding the rotation, and to provide a good optical quality. The goal of this study was to investigate the influence of the material and haptic design on the behavior of the IOLs under dynamic compression condition. For this purpose, the strain–stress characteristics of the hydrophobic and hydrophilic materials were estimated experimentally. Next, these data were used as the input for a finite-element model (FEM) to analyze the stability of different IOL haptic designs, according to the procedure described by the ISO standards. Finally, the simulations of the effect of IOL tilt and decentration on the optical performance were performed in an eye model using a ray-tracing software. The results suggest the major importance of the haptic design rather than the material on the postoperative behavior of an IOL. FEM appears to be a powerful tool for numerical studies of the biomechanical properties of IOLs and it allows one to help in the design phase to the manufacturers.

This study is focused on the measurements of the refractive index of hemoglobin solutions in the visible/near-infrared (NIR) spectral range at room temperature for characteristic laser wavelengths: 480, 486, 546, 589, 644, 656, 680, 930, 1100, 1300, and 1550 nm. Measurements were performed using the multiwavelength Abbe refractometer. Aqua hemoglobin solutions of different concentrations obtained from human whole blood were investigated. The specific increment of refractive index on hemoglobin concentration and the Sellmeier coefficients were calculated.

The present research intends to propose a fully automatic algorithm for the classification of three-dimensional (3-D) optical coherence tomography (OCT) scans of patients suffering from abnormal macula from normal candidates. The method proposed does not require any denoising, segmentation, retinal alignment processes to assess the intraretinal layers, as well as abnormalities or lesion structures. To classify abnormal cases from the control group, a two-stage scheme was utilized, which consists of automatic subsystems for adaptive feature learning and diagnostic scoring. In the first stage, a wavelet-based convolutional neural network (CNN) model was introduced and exploited to generate B-scan representative CNN codes in the spatial-frequency domain, and the cumulative features of 3-D volumes were extracted. In the second stage, the presence of abnormalities in 3-D OCTs was scored over the extracted features. Two different retinal SD-OCT datasets are used for evaluation of the algorithm based on the unbiased fivefold cross-validation (CV) approach. The first set constitutes 3-D OCT images of 30 normal subjects and 30 diabetic macular edema (DME) patients captured from the Topcon device. The second publicly available set consists of 45 subjects with a distribution of 15 patients in age-related macular degeneration, DME, and normal classes from the Heidelberg device. With the application of the algorithm on overall OCT volumes and 10 repetitions of the fivefold CV, the proposed scheme obtained an average precision of 99.33% on dataset1 as a two-class classification problem and 98.67% on dataset2 as a three-class classification task.

Nicotinamide has been shown to affect blood flow in both tumor and normal tissues, including skeletal muscle. Intraperitoneal injection of nicotinamide was used as a simple intervention to test the sensitivity of noninvasive diffuse correlation spectroscopy (DCS) to changes in blood flow in the murine left quadriceps femoris skeletal muscle. DCS was then compared with the gold-standard fluorescent microsphere (FM) technique for validation. The nicotinamide dose-response experiment showed that relative blood flow measured by DCS increased following treatment with 500- and 1000-mg  /  kg nicotinamide. The DCS and FM technique comparison showed that blood flow index measured by DCS was correlated with FM counts quantified by image analysis. The results of this study show that DCS is sensitive to nicotinamide-induced blood flow elevation in the murine left quadriceps femoris. Additionally, the results of the comparison were consistent with similar studies in higher-order animal models, suggesting that mouse models can be effectively employed to investigate the utility of DCS for various blood flow measurement applications.

A practical algorithm for estimating the wavelength-dependent refractive index (RI) of a turbid sample in the spatial frequency domain with the aid of Kramers–Kronig (KK) relations is presented. In it, phase-shifted sinusoidal patterns (structured illumination) are serially projected at a high spatial frequency onto the sample surface (mouse scalp) at different near-infrared wavelengths while a camera mounted normally to the sample surface captures the reflected diffuse light. In the offline analysis pipeline, recorded images at each wavelength are converted to spatial absorption maps by logarithmic function, and once the absorption coefficient information is obtained, the imaginary part (k) of the complex RI (CRI), based on Maxell’s equations, can be calculated. Using the data represented by k, the real part of the CRI (n) is then resolved by KK analysis. The wavelength dependence of n  (  λ  )   is then fitted separately using four standard dispersion models: Cornu, Cauchy, Conrady, and Sellmeier. In addition, three-dimensional surface-profile distribution of n is provided based on phase profilometry principles and a phase-unwrapping-based phase-derivative-variance algorithm. Experimental results demonstrate the capability of the proposed idea for sample’s determination of a biological sample’s RI value.

Marginal gap (MG) was the most important factor to evaluate the success of crowns. The study was to assess the MG of lithium disilicate glass ceramic crowns with spectral domain optical coherence tomography (OCT) and stereomicroscopy in vitro, and to provide evidence to measure the MG with OCT. Consistency was observed between OCT and stereomicroscopy to measure the MG after cementation. There was no significant difference between the MG of lithium disilicate glass ceramic crowns using OCT and stereomicroscopy (OCT 59.55  ±  7.22  μm, stereomicroscope 59.48  ±  6.53  μm, P  =  0.736) after cementation. OCT was a noninvasive diagnostic technique to measure the MG of lithium disilicate glass ceramic crowns.

The diagnosis of corneal diseases may be improved by monitoring the metabolism of cells and the structural organization of the stroma using two-photon imaging (TPI). We used TPI to assess the differences between nonpathological (NP) human corneas and corneas diagnosed with either keratoconus, Acanthamoeba keratitis, or stromal corneal scars. Images were acquired using a custom-built five-dimensional laser-scanning microscope with a broadband sub-15 femtosecond near-infrared pulsed excitation laser and a 16-channel photomultiplier tube detector in combination with a time-correlated single photon counting module. Morphological alterations of epithelial cells were observed for all pathologies. Moreover, diseased corneas showed alterations to the cells’ metabolism that were revealed using the NAD(P)H free to protein-bound ratios. The mean autofluorescence lifetime of the stroma and the organization of the collagen fibers were also significantly altered due to the pathologies. We demonstrate that TPI can be used to distinguish between NP and diseased human corneas, based not only on alterations of the cells’ morphology, which can also be evaluated using current clinical devices, but on additional morphological and functional features such as the organization of the stroma and the cells’ metabolism. Therefore, TPI could become an efficient tool for diagnosing corneal diseases and better understanding the biological processes of the diseases.

We have designed and implemented a dual-mode adaptive optics (AO) imaging system that combines spectral domain optical coherence tomography (OCT) and scanning laser ophthalmoscopy (SLO) for in vivo imaging of the human retina. The system simultaneously acquires SLO frames and OCT B-scans at 60 Hz with an OCT volume acquisition time of 4.2 s. Transverse eye motion measured from the SLO is used to register the OCT B-scans to generate three-dimensional (3-D) volumes. Key optical design considerations include: minimizing system aberrations through the use of off-axis relay telescopes, conjugate pupil plane requirements, and the use of dichroic beam splitters to separate and recombine the OCT and SLO beams around the nonshared horizontal scanning mirrors. To demonstrate system performance, AO-OCT-SLO images and measurements are taken from three normal human subjects ranging in retinal eccentricity from the fovea out to 15-deg temporal and 20-deg superior. Also presented are en face OCT projections generated from the registered 3-D volumes. The ability to acquire high-resolution 3-D images of the human retina in the midperiphery and beyond has clinical importance in diseases, such as retinitis pigmentosa and cone–rod dystrophy.

We investigate the use of a fiber-based, multispectral fluorescence lifetime imaging (FLIm) system to nondestructively monitor changes in mechanical properties of collagen hydrogels caused by controlled application of widely used cross-linking agents, glutaraldehyde (GTA) and ribose. Postcross-linking, fluorescence lifetime images are acquired prior to the hydrogels being processed by rheological or tensile testing to directly probe gel mechanical properties. To preserve the sterility of the ribose-treated gels, FLIm is performed inside a biosafety cabinet (BSC). A pairwise correlation analysis is used to quantify the relationship between mean hydrogel fluorescence lifetimes and the storage or Young’s moduli of the gels. In the GTA study, we observe strong and specific correlations between fluorescence lifetime and the storage and Young’s moduli. Similar correlations are not observed in the ribose study and we postulate a reason for this. Finally, we demonstrate the ability of FLIm to longitudinally monitor dynamic cross-link formation. The strength of the GTA correlations and deployment of our fiber-based FLIm system inside the aseptic environment of a BSC suggests that this technique may be a valuable tool for the tissue engineering community where longitudinal assessment of tissue construct maturation in vitro is highly desirable.

Chorioretinal imaging has a crucial role for the patients with chorioretinal vascular diseases, such as neovascular age-related macular degeneration. Imaging oxygen gradients in the eye could better diagnose and treat ocular diseases. Here, we describe the use of photoacoustic ocular imaging (PAOI) in measuring chorioretinal oxygen saturation (CR  −  sO₂) gradients in New Zealand white rabbits (n  =  5) with ocular ischemia. We observed good correlation (R²  =  0.98) between pulse oximetry and PAOI as a function of different oxygen percentages in inhaled air. We then used an established ocular ischemia model in which intraocular pressure is elevated to constrict ocular blood flow, and notice a positive correlation (R²  =  0.92) between the injected volume of phosphate buffered saline (PBS) and intraocular pressure (IOP) as well as a negative correlation (R²  =  0.98) between CR  −  sO₂ and injected volume of PBS. The CR  −  sO₂ was measured before (baseline), during (ischemia), and after the infusion (600-μL PBS). The ischemia-reperfusion model did not affect the measurement of the sO₂ using a pulse oximeter on the animal’s paw, but the chorioretinal PAOI signal showed a nearly sixfold decrease in CR  −  sO₂ (n  =  5, p  =  0.00001). We also observe a sixfold decrease in CR  −  sO₂ after significant elevation of IOP during ischemia, with an increase close to baseline during reperfusion. These data suggest that PAOI can detect changes in chorioretinal oxygenation and may be useful for application to imaging oxygen gradients in ocular disease.

Laser speckle imaging (LSI) is a wide-field optical technique that enables superficial blood flow quantification. LSI is normally performed in a mounted configuration to decrease the likelihood of motion artifact. However, mounted LSI systems are cumbersome and difficult to transport quickly in a clinical setting for which portability is essential in providing bedside patient care. To address this issue, we created a handheld LSI device using scientific grade components. To account for motion artifact of the LSI device used in a handheld setup, we incorporated a fiducial marker (FM) into our imaging protocol and determined the difference between highest and lowest speckle contrast values for the FM within each data set (K_best and K_worst). The difference between K_best and K_worst in mounted and handheld setups was 8% and 52%, respectively, thereby reinforcing the need for motion artifact quantification. When using a threshold FM speckle contrast value (K_FM) to identify a subset of images with an acceptable level of motion artifact, mounted and handheld LSI measurements of speckle contrast of a flow region (K_FLOW) in in vitro flow phantom experiments differed by 8%. Without the use of the FM, mounted and handheld K_FLOW values differed by 20%. To further validate our handheld LSI device, we compared mounted and handheld data from an in vivo porcine burn model of superficial and full thickness burns. The speckle contrast within the burn region (K_BURN) of the mounted and handheld LSI data differed by <4 %   when accounting for motion artifact using the FM, which is less than the speckle contrast difference between superficial and full thickness burns. Collectively, our results suggest the potential of handheld LSI with an FM as a suitable alternative to mounted LSI, especially in challenging clinical settings with space limitations such as the intensive care unit.

Ultraviolet photoacoustic microscopy (UV-PAM) is a promising intraoperative tool for surgical margin assessment (SMA), one that can provide label-free histology-like images with high resolution. In this study, using a microlens array and a one-dimensional (1-D) array ultrasonic transducer, we developed a high-throughput multifocal UV-PAM (MF-UV-PAM). Our new system achieved a 1.6  ±  0.2  μm lateral resolution and produced images 40 times faster than the previously developed point-by-point scanning UV-PAM. MF-UV-PAM provided a readily comprehensible photoacoustic image of a mouse brain slice with specific absorption contrast in ∼16  min, highlighting cell nuclei. Individual cell nuclei could be clearly resolved, showing its practical potential for intraoperative SMA.

Line-scan focal modulation microscopy (LSFMM) is an emerging imaging technique that affords high imaging speed and good optical sectioning at the same time. We present a systematic investigation into optimal design of the pupil filter for LSFMM in an attempt to achieve the best performance in terms of spatial resolutions, optical sectioning, and modulation depth. Scalar diffraction theory was used to compute light propagation and distribution in the system and theoretical predictions on system performance, which were then compared with experimental results.

A protocol for photoacoustic imaging (PAI) has been developed to assess pixel-based oxygen saturation (sO₂) distributions of experimental tumor models. The protocol was applied to evaluate the dependence of PAI results on measurement settings, reproducibility of PAI, and for the characterization of the oxygenation status of experimental prostate tumor sublines (Dunning R3327-H, -HI, -AT1) implanted subcutaneously in male Copenhagen rats. The three-dimensional (3-D) PA data employing two wavelengths were used to estimate sO₂ distributions. If the PA signal was sufficiently strong, the distributions were independent from signal gain, threshold, and positioning of animals. Reproducibility of sO₂ distributions with respect to shape and median values was demonstrated over several days. The three tumor sublines were characterized by the shapes of their sO₂ distributions and their temporal response after external changes of the oxygen supply (100% O₂ or air breathing and clamping of tumor-supplying artery). The established protocol showed to be suitable for detecting temporal changes in tumor oxygenation as well as differences in oxygenation between tumor sublines. PA results were in accordance with histology for hypoxia, perfusion, and vasculature. The presented protocol for the assessment of pixel-based sO₂ distributions provides more detailed information as compared to conventional region-of-interest-based analysis of PAI, especially with respect to the detection of temporal changes and tumor heterogeneity.

Polymeric endovascular implants are the next step in minimally invasive vascular interventions. As an alternative to traditional metallic drug-eluting stents, these often-erodible scaffolds present opportunities and challenges for patients and clinicians. Theoretically, as they resorb and are absorbed over time, they obviate the long-term complications of permanent implants, but in the short-term visualization and therefore positioning is problematic. Polymeric scaffolds can only be fully imaged using optical coherence tomography (OCT) imaging—they are relatively invisible via angiography—and segmentation of polymeric struts in OCT images is performed manually, a laborious and intractable procedure for large datasets. Traditional lumen detection methods using implant struts as boundary limits fail in images with polymeric implants. Therefore, it is necessary to develop an automated method to detect polymeric struts and luminal borders in OCT images; we present such a fully automated algorithm. Accuracy was validated using expert annotations on 1140 OCT images with a positive predictive value of 0.93 for strut detection and an R2 correlation coefficient of 0.94 between detected and expert-annotated lumen areas. The proposed algorithm allows for rapid, accurate, and automated detection of polymeric struts and the luminal border in OCT images.

We present a nonlocal weighted sparse representation (NWSR) method for reconstruction of retinal optical coherence tomography (OCT) images. To reconstruct a high signal-to-noise ratio and high-resolution OCT images, utilization of efficient denoising and interpolation algorithms are necessary, especially when the original data were subsampled during acquisition. However, the OCT images suffer from the presence of a high level of noise, which makes the estimation of sparse representations a difficult task. Thus, the proposed NWSR method merges sparse representations of multiple similar noisy and denoised patches to better estimate a sparse representation for each patch. First, the sparse representation of each patch is independently computed over an overcomplete dictionary, and then a nonlocal weighted sparse coefficient is computed by averaging representations of similar patches. Since the sparsity can reveal relevant information from noisy patches, combining noisy and denoised patches’ representations is beneficial to obtain a more robust estimate of the unknown sparse representation. The denoised patches are obtained by applying an off-the-shelf image denoising method and our method provides an efficient way to exploit information from noisy and denoised patches’ representations. The experimental results on denoising and interpolation of spectral domain OCT images demonstrated the effectiveness of the proposed NWSR method over existing state-of-the-art methods.

Visible light optical coherence tomography has shown great interest in recent years for spectroscopic and high-resolution retinal and cerebral imaging. Here, we present an extended-focus optical coherence microscopy system operating from the visible to the near-infrared wavelength range for high axial and lateral resolution imaging of cortical structures in vivo. The system exploits an ultrabroad illumination spectrum centered in the visible wavelength range (λc  =  650  nm, Δλ  ∼  250  nm) offering a submicron axial resolution (∼0.85  μm in water) and an extended-focus configuration providing a high lateral resolution of ∼1.4  μm maintained over ∼150  μm in depth in water. The system’s axial and lateral resolution are first characterized using phantoms, and its imaging performance is then demonstrated by imaging the vasculature, myelinated axons, and neuronal cells in the first layers of the somatosensory cortex of mice in vivo.

The tissue metabolic rate of oxygen consumption (tMRO2) is a clinically relevant marker for a number of pathologies including cancer and arterial occlusive disease. We present and validate a noncontact method for quantitatively mapping tMRO2 over a wide, scalable field of view at 16  frames  /  s. We achieve this by developing a dual-wavelength, near-infrared coherent spatial frequency-domain imaging (cSFDI) system to calculate tissue optical properties (i.e., absorption, μa, and reduced scattering, μs′, parameters) as well as the speckle flow index (SFI) at every pixel. Images of tissue oxy- and deoxyhemoglobin concentration (  [  HbO2  ]   and [HHb]) are calculated from optical properties and combined with SFI to calculate tMRO2. We validate the system using a series of yeast-hemoglobin tissue-simulating phantoms and conduct in vivo tests in humans using arterial occlusions that demonstrate sensitivity to tissue metabolic oxygen debt and its repayment. Finally, we image the impact of cyanide exposure and toxicity reversal in an in vivo rabbit model showing clear instances of mitochondrial uncoupling and significantly diminished tMRO2. We conclude that dual-wavelength cSFDI provides rapid, quantitative, wide-field mapping of tMRO2 that can reveal unique spatial and temporal dynamics relevant to tissue pathology and viability.

As a high-resolution imaging mode of biological tissues and materials, optical coherence tomography (OCT) is widely used in medical diagnosis and analysis. However, OCT images are often degraded by annoying speckle noise inherent in its imaging process. Employing the bilateral sparse representation an adaptive singular value shrinking method is proposed for its highly sparse approximation of image data. Adopting the generalized likelihood ratio as similarity criterion for block matching and an adaptive feature-oriented backward projection strategy, the proposed algorithm can restore better underlying layered structures and details of the OCT image with effective speckle attenuation. The experimental results demonstrate that the proposed algorithm achieves a state-of-the-art despeckling performance in terms of both quantitative measurement and visual interpretation.

We demonstrated that different degrees of experimental traumatic brain injury (TBI) can be differentiated clearly in fresh slices of rat brain tissues using transmission-type terahertz (THz) imaging system. The high absorption region in THz images corresponded well with the injured area in visible images and magnetic resonance imaging results. The THz image and absorption characteristics of dehydrated paraffin-embedded brain slices and the hematoxylin and eosin (H&amp;E)-stained microscopic images were investigated to account for the intrinsic differences in the THz images for the brain tissues suffered from different degrees of TBI and normal tissue aside from water. The THz absorption coefficients of rat brain tissues showed an increase in the aggravation of brain damage, particularly in the high-frequency range, whereas the cell density decreased as the order of mild, moderate, and severe TBI tissues compared with the normal tissue. Our results indicated that the different degrees of TBI were distinguishable owing to the different water contents and probable hematoma components distribution rather than intrinsic cell intensity. These promising results suggest that THz imaging has great potential as an alternative method for the fast diagnosis of TBI.

Effectiveness of extended depth of field microscopy (EDFM) implementation with wavefront encoding methods is reduced by depth-induced spherical aberration (SA) due to reliance of this approach on a defined point spread function (PSF). Evaluation of the engineered PSF’s robustness to SA, when a specific phase mask design is used, is presented in terms of the final restored image quality. Synthetic intermediate images were generated using selected generalized cubic and cubic phase mask designs. Experimental intermediate images were acquired using the same phase mask designs projected from a liquid crystal spatial light modulator. Intermediate images were restored using the penalized space-invariant expectation maximization and the regularized linear least squares algorithms. In the presence of depth-induced SA, systems characterized by radially symmetric PSFs, coupled with model-based computational methods, achieve microscope imaging performance with fewer deviations in structural fidelity (e.g., artifacts) in simulation and experiment and 50% more accurate positioning of 1-μm beads at 10-μm depth in simulation than those with radially asymmetric PSFs. Despite a drop in the signal-to-noise ratio after processing, EDFM is shown to achieve the conventional resolution limit when a model-based reconstruction algorithm with appropriate regularization is used. These trends are also found in images of fixed fluorescently labeled brine shrimp, not adjacent to the coverslip, and fluorescently labeled mitochondria in live cells.

Choosing the localization algorithm in superresolution microscopy is an important factor in determining the resolution in such a modality. The point spread function (PSF) in defocused images has ring structures that can be used to localize the three-dimensional (3-D) position of single particles by calculating the ring center (x and y) and radius (z). As there is no well-developed mathematical model for a defocused PSF, it is difficult to perform a fitting-based algorithm in such images. A particle localization algorithm based on radial symmetry and ellipse fitting is developed to localize the centers and radii of defocused PSFs. Our method can localize the 3-D position of a fluorophore within 20-nm precision in three-dimensions in a range of 40  μm in z-dimension from defocused two-dimensional (2-D) images.

Active infrared thermography is a fast, painless, noncontact, and noninvasive imaging method, complementary to mammography, ultrasound, and magnetic resonance imaging methods for early diagnosis of breast cancer. This technique plays an important role in early detection of breast cancer to women of all ages, including pregnant or nursing women, with different sizes of breast, irrespective of either fatty or dense breast. This proposed complementary technique makes use of infrared emission emanating from the breast. Emanating radiations from the surface of the breast under test are detected with an infrared camera to map the thermal gradients over it, in order to reveal hidden tumors inside it. One of the reliable active infrared thermographic technique, linear frequency modulated thermal wave imaging is adopted to detect tumors present inside the breast. Further, phase and amplitude images are constructed using frequency and time-domain data analysis schemes. Obtained results show the potential of the proposed technique for early diagnosis of breast cancer in fatty as well as dense breasts.